无机焦磷酸酶GMP级|Pyrophosphatase,Inorganic GMP-grade

发表于2024年10月25日由上海金畔生物科技有限公司

产品说明书

FAQ

COA

已发表文献

Pyrophosphatase, Inorganic GMP-grade (1 U/μL) is an inorganic pyrophosphatase derived from recombinant expression in E. coli. It is an enzyme that can catalyze the conversion of one molecule of pyrophosphate into two molecules of phosphate ions. This is a high-energy reaction, so this reaction can be coupled to some thermodynamically unfavorable transformations in order to drive these transformations to completion. Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to orthophosphate. In molecular biology, it can be used to increase RNA yield in reverse transcription reaction.

This product is produced in accordance with GMP process requirements, and the product is provided in liquid form

Product Properties

Source	Recombinant E.coli with Pyrophosphatase gene from yeast
Optimum Temperature	25℃
Storage Buffer	50 mM Tris-HCl pH 7.9, 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% （v/v）TritonX-100, 50%（v/v）glycerin
Unit Definition	One unit (U) is the amount of enzyme needed to catalyze the hydrolysis of PPI per minute to produce 1 μmol Pi.（The standard reaction is: 500 μL system contains 100 mM Tris-HCl pH7.2, 2 mM MgCl₂, 2 mM Ppi; reaction at 25℃ for 10 min）

Contents

Catalog No.

Name

Catalog No./Specification

10620ES10

(10 U)

10620ES60

(100 U)

10620ES80

(1,000 U)

10620ES99

(40 KU)

10620

Pyrophosphatase, Inorganic GMP-grade (1 U/μL)

10 μL

100 μL

1 mL

40 mL

Shipping and Storage

The Pyrophosphatase, Inorganic GMP-grade products shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. This product is active in a variety of reaction buffers. Usually, this product can be directly connected in experiments such as IVT.

2.The optimal reaction temperature of this product is 25℃, it is active at 16-37℃, 65℃ can inactivate the enzyme for 10 minutes.

3. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this.

HB220701

This product is produced in accordance with GMP process requirements, and the product is provided in liquid form

Product Properties

Source	Recombinant E.coli with Pyrophosphatase gene from yeast
Optimum Temperature	25℃
Storage Buffer	50 mM Tris-HCl pH 7.9, 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% （v/v）TritonX-100, 50%（v/v）glycerin
Unit Definition	One unit (U) is the amount of enzyme needed to catalyze the hydrolysis of PPI per minute to produce 1 μmol Pi.（The standard reaction is: 500 μL system contains 100 mM Tris-HCl pH7.2, 2 mM MgCl₂, 2 mM Ppi; reaction at 25℃ for 10 min）

Contents

Catalog No.

Name

Catalog No./Specification

10620ES10

(10 U)

10620ES60

(100 U)

10620ES80

(1,000 U)

10620ES99

(40 KU)

10620

Pyrophosphatase, Inorganic GMP-grade (1 U/μL)

10 μL

100 μL

1 mL

40 mL

Shipping and Storage

The Pyrophosphatase, Inorganic GMP-grade products shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. This product is active in a variety of reaction buffers. Usually, this product can be directly connected in experiments such as IVT.

2.The optimal reaction temperature of this product is 25℃, it is active at 16-37℃, 65℃ can inactivate the enzyme for 10 minutes.

3. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this.

HB220701

无机焦磷酸酶GMP级|Pyrophosphatase,Inorganic GMP-grade

暂无内容

[1] Li Q, You J, Qiao T, Zhong DB, Yu X. Sodium chloride stimulates the biomass and astaxanthin production by Haematococcus pluvialis via a two-stage cultivation strategy. Bioresour Technol. 2022;344(Pt A):126214. doi:10.1016/j.biortech.2021.126214(IF:9.642)
[2] Ma Y, Cheng B, Li Y, et al. Protective Effect of Nanoselenium on Renal Oxidative Damage Induced by Mercury in Laying Hens. Biol Trace Elem Res. 2022;200(8):3785-3797. doi:10.1007/s12011-021-02956-z(IF:3.738)

GMP级干细胞冻存液

发表于2024年10月17日由上海金畔生物科技有限公司

上海金畔生物科技有限公司amsbio专业代理，AMSBIO成立于1987年，公司由艾伦和亚历克斯联合创立，总部位于西班牙马德里，AMSBIO在意大利，瑞士和英国均有办事处。AMSBIO的使命是成为一个生命科学研究的试剂和服务帮助客户开发创新的方法，过程，产品和药品的首要供应商，为学术团体和生物技术合作伙伴提供的技术和成本有效的解决方案。

Amsbio STEM-CELLBANKER® 说明书

品牌：Amsbio

货号： 11897

品名： STEM-CELLBANKER

描述：GMP Grade Stem Cell Cryopreservation Media

产品信息

产品名称	产品编号	规格	价格（元）
STEM-CELLBANKER ® – GMP	11890	100ml	3700.00
STEM-CELLBANKER ® – GMP	11897	20ml	1300
STEM-CELLBANKER ® – GMP 不含DMSO	11897F	20ml	1300
STEM-CELLBANKER ® – GMP 不含DMSO	11890F	100ml	3700

产品描述

STEM-CELLBANKER ^®是GMP下生产的ES细胞和iPS细胞冻存液产品特点是化学成分清晰，无动物源成分，专门针对ES细胞和iPS细胞特点设计的细胞冻存液

· 化学成分清楚和无动物源成分

· 按照日本，欧盟，美国和PIC / S GMP指南生产

· 复苏后保持细胞的高活力

· 解冻复苏后依然保持干细胞的多能性

· 实现细胞的长期冻存，不管是-80度还是液氮保存都可以保证活性10年以上

· 可以不需要液氮就可以长期保存

· 同样可以适合组织冷冻保存及其他珍贵细胞冻存

· STEM-CELLBANKER 已经得到美国食品药物管理局的认证（生物制品评价和研究中心）

STEM-CELLBANKER ^®是*无的血清和动物源性成分的，所有成分*欧洲药典或美国药典的要求，是干细胞治疗和应用zui合适的细胞冻存液，STEM-CELLBANKER ^®使用方便，不需要特殊设备和仪器就可以实现干细胞高存活率的冻存和长达10年以上的长期保存。

STEM-CELLBANKER ^®^还显著增加细胞活力，同时保持细胞多能性，和细胞的正常核型和增殖能力，是一般冻存液无法达到的。

应用实例

使用STEM-CELLBANKER对人体胚胎干细胞（hESC） 和诱导多能（iPS）干细胞进行实际冻存测试。

测试冻存液：STEM-CELLBANKER冻存液 vs 常规细胞冻存液（细胞培养基 + 10% DMSO + 血清代替品）
细胞存活率评估方法：trypan Blue 和 Live/Dead assay kit试剂盒

测试结果：使用STEM-CELLBANKER冻存液的细胞存活率明显提高，HS293, HS306 and ChiPSA 分别高达９３%、９６%　和９０％，比只有５０％存活率的常规细胞冻存液高一倍。同时，结果也显示STEM-CELLBANKER冻存液没有给细胞的增殖、分化和染色体组形带来影响。

STEM-CELLBANKER用于小鼠肾脏组织冻存比较图：

图2.将小鼠肾脏切碎成约5平方毫米。将组织样品或者立即固定用10％福尔马林或冷冻保存与在-801毫升STEM-CELLBANKER的^ø C一周。冷冻保存的样品在解冻后立即用10％福尔马林固定。两种样品的石蜡切片用H＆E或抗活性半胱天冬酶3抗体染色。在新鲜和冷冻保存的组织样品中，坏死或细胞凋亡的迹象都可以忽略不计。在来自各种小鼠器官的组织样品的情况下获得了类似的结果。

STEM-CELLBANKER保存的细胞MRNA保存水平图

图3.总RNA从小鼠肺成纤维细胞系的任收获后立即或在-80与STEM-CELLBANKER冻存后的细胞中提取^ø C一周和解冻。通过DNA微阵列分析比较两个样品中的mRNA水平，结果显示93％的mRNA在±1.6倍差异的范围内。

使用STEM-CELLBANKER小鼠胚胎干细胞（129SV）进行实际冻存测试。

细胞株：129SV (Mouse ES cell)
测试冻存液：ZENOAQ 产品（CELLBANKER 1, 1plus, 2 和 STEM-CELLBANKER）
常规自培冻存液（90% FBS+10%DMSO 和 90% culture medium + 10%DMSO

操作流程图：

使用手册:

细胞冷冻保存方法

选择冻存处于对数生长数的细胞有助于提高复苏细胞存活率。

1. 按照常用方法收集悬浮细胞或贴壁细胞于试管中。

2. 按照培养细胞密度和所用细胞冻存管的尺寸计算所需冻存细胞数。
（参考：5×105 至 5×106 cells/ml）。

3. 取相当于所需细胞数的细胞悬浮液量，置于离心管中，离心收集培养细胞(参考离心条件：1,000~2,000rpm，4℃，3至5分)。移去离心管中的上清液。

4. 加入适量的CELLBANKERⓇ 细胞冻存液于离心管中，使细胞浓度为 5×105 至 5×106 cells/ml。缓慢地混合均匀，制成细胞混合液。

5. 将离心管中的细胞混合液分装于已标示*的冷冻保存管中。

6. 直接将含细胞混合液的冻存管放入-80℃冰箱，长期冷冻保存。

7. 若研究者需要液氮保存时，可将*冻结的冻存管（放入-80℃冰箱后至少一昼夜）移至-196℃液氮罐。

冻存细胞复苏方法

1. 从冰箱里取出细胞冷冻保存管，立即放入37℃ 振动水浴槽中快速解冻。

2. 待冻存管中细胞混合液*融化后，立即加入1ml细胞培养基于该冷冻管中与细胞混合，再将细胞混合液从冻存管中移入含有9ml该细胞培养基的试管中，混合均匀。

3. 离心收集培养细胞((参考离心条件：1,000~2,000rpm，4℃，3至5分)，移去上清液。

4. 清洗细胞，充分洗净残留冻存液。

5. 加入适量的新鲜细胞培养基，使用移液管缓缓地均匀细胞混合液。适量地稀释后，将细胞混合液移至事先准备好的培养瓶中。

6. 精检后，研究者可根据各自方法和需要来进行细胞培养。

储存方法和保质期

储存于４度以下。保质期３年。3个月以上没有使用CELLBANKER冻存液的计划时，尽可能冷冻保存。为避免重复冻结–解冻过程可能导致的CELLBANKER品质下降和性能降低，推荐将 100mL 产品分装小瓶后，再存放于 4℃以下或－20℃冰箱冻存。

质量保障承诺

1. STEM-CELLBANKER 细胞冻存液经过严格内毒素、pH、渗透压和微生物检查，确保产品不含细菌、霉菌以及支原体。

2. 使用小鼠胚胎干细胞（Mouse ES Cells）、Jurkat　和SK-007细胞验证性能测试冻存细保存活率达８０％以上．

TESTS	SPECIFICATION
Endotoxin（内毒素）	低于 20 EU/ml
Mycoplasma（支原体）	无
pH	7.0 – 8.5
Osmolarity（容量渗透压）	低于1800 mOsm
Sterility (微生物检查)	无

注意事项

1. CELLBANKER系列产品只供实验室研究使用，是试剂级产品。

2. 我们推荐用户在使用本产品之前，事先对所使用的细胞进行至少为期 1 周的
试验性细胞冷冻保存培养。确认性能后再进行冻存。

3. 免责声明: 本公司将不为任何不正常使用此产品时所发生的意外负责。

amsbio 其它产品信息：

货号	品名	中文名称	规格	目录价	品牌
11891	Cellbanker2	无血清型细胞冻存液	100ml	1580	amsbio
11892	Cellbanker2	无血清型细胞冻存液	20ml	420	amsbio
11890	Stem-Cellbanker GMP grade	GMP级干细胞冻存液	100ml	3500	amsbio
11897	Stem-Cellbanker GMP grade	GMP级干细胞冻存液	20ml	838	amsbio
11890F	Stem-Cellbanker DMSO Free GMP grade	无DMSO干细胞冻存液	100ml	3650	amsbio
11897F	Stem-Cellbanker DMSO Free GMP grade	无DMSO干细胞冻存液	20ml	838	amsbio
11898	Cellotion	无血清型细胞清洗液	100ml	768	amsbio

上海金畔生物科技有限公司是实验试剂一站式采购服务商

1：强大的进口辐射能力，血清、抗体、耗材、大部分限制进口品等。

2：产品种类齐全，经营超过700多个品牌，基本涵盖所有生物实验试剂耗材。

3：提供加急服务，货品一般1-2周到货。

4：富有竞争力的价格优势，绝大部分价格有优势。

5：多年积累良好的信誉，大部分客户提供货到付款服务。客户包括清华、北大、交大、复旦、中山等100多所高校，ROCHE，阿斯利康、国药、fisher等药企。

6：我们还是Santa,Advanced Biotechnologies Inc,Athens Research & Technology,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. FormuMax Scientific,Inc, Genebridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,omicronbio, Polysciences,prospecbi, QA-BIO,quickzyme,RESEARCH DIETS,INC,sterlitech；sysy,TriLink BioTechnologies,Inc；worthington-biochem,zyagen等几十家国外公司代理。

7：我们还是invitrogen,qiagen,MiraiBioam,sigma;neb,roche,merck, rnd,BD, GE,pierce,BioLegend等*批发，欢迎合作。

CTP Tris溶液GMP级(100 mM) CTP Tris Solution

发表于2024年10月11日由上海金畔生物科技有限公司

CTP Tris溶液GMP级(100 mM) CTP Tris Solution

产品说明书

FAQ

COA

已发表文献

Product Information

Product name	Cat#	Size
CTP Tris Solution GMP-grade (100 mM)	10653ES03	1 mL
	10653ES10	10 mL
	10653ES60	100 mL

Product Description

CTP, Cytidine-5'-triphosphate, can be used in a variety of molecular biology applications, such as in vitro transcription, RNA amplification, and the synthesis of choline and ethanolamine.

This product is a transparent colorless aqueous solution prepared from CTP tris solution, and free from DNase, RNase and phosphatase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	65-47-4 (free acid)
Formula	C₉H₁₆N₃O₁₄P₃(free acid)
Molecular weight	483.16 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

Product Information

Product name	Cat#	Size
CTP Tris Solution GMP-grade (100 mM)	10653ES03	1 mL
	10653ES10	10 mL
	10653ES60	100 mL

Product Description

CTP, Cytidine-5'-triphosphate, can be used in a variety of molecular biology applications, such as in vitro transcription, RNA amplification, and the synthesis of choline and ethanolamine.

This product is a transparent colorless aqueous solution prepared from CTP tris solution, and free from DNase, RNase and phosphatase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	65-47-4 (free acid)
Formula	C₉H₁₆N₃O₁₄P₃(free acid)
Molecular weight	483.16 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

CTP Tris溶液GMP级(100 mM) CTP Tris Solution

暂无内容

PeproTech GMP120-14E说明书

发表于2024年10月2日由上海金畔生物科技有限公司

随着细胞疗法从实验室过渡到患者的床边，确保高水平的质量和安全性是一项重大而关键的挑战。PeproTech公司的PeproGMP ®细胞因子帮助研究人员应对这一挑战。与联邦要求和细胞治疗，基因治疗和组织工程产品，为临床应用的宜实践一致的，PeproGMP ®细胞因子生产并符合美国FDA的GMP（良好生产规范）的规定和ISO 9001质量管理体系标准进行测试，不使用动物源性材料。

PeproGMP ®细胞因子提供安全性，纯度，和在离体的制造工艺简化的用途，作为用于细胞，基因，和组织工程产品在USP（美国药典）第<1043>辅助材料进行说明。

所有PeproGMP®产品均采用动物源性无组分材料制造。

PeproTech GMP120-14E说明书

PeproGMP®重组人激活素A.

PeproGMP® Recombinant Human Activin A

Product Details

产品详情

目录号：GMP120-14E

规格：

50μg

100μG

描述：

激活素A是TGF-β家族成员，具有广泛的生物活性，包括细胞增殖和分化的调节，以及促进神经元存活。

• 分析证书上提供了其他产品信息和规格。

注意：所有PeproGMP®产品均采用动物源性无组分材料制造。

AA序列（单体）：GLECDGKVNI CCKKQFFVSF KDIGWNDWII APSGYHANYC EGECPSHIAG TSGSSLSFHS TVINHYRMRG HSPFANLKSC CVPTKLRPMS MLYYDDGQNI IKKDIQNMIV EECGCS

纯度：通过SDS-PAGE凝胶和HPLC分析≥98％。

来源：大肠杆菌

原产地：美国

不适合人类使用。

mRNA Cap 2氧甲基转移酶GMP级|mRNA Cap 2´-O-Methyltransferase GMP-grade

发表于2024年9月28日由上海金畔生物科技有限公司

mRNA Cap 2氧甲基转移酶GMP级|mRNA Cap 2´-O-Methyltransferase GMP-grade

产品说明书

FAQ

COA

已发表文献

Product Description

mRNA Cap 2´-O-Methyltransferase is a recombinant protein derived from vaccinia virus. The enzyme can add a methyl group at the 2´-O position of the first nucleotide of the cap structure at the 5´ end of the RNA. The enzyme uses SAM as a methyl donor to methylate capped RNA to form a Cap1 structure. The Cap1 structure can enhance the translation efficiency of mRNA, so it can improve the expression of mRNA in transfection and microinjection experiments. This enzyme requires RNA with m7GpppN, a 7-methylguanosine cap structure, as a substrate.

This product is produced in accordance with GMP process requirements. The product is provided in liquid form and can be used for Cap1 capping reaction of pre-mRNA in vivo/in vitro.

Product Properties

Source	Recombinant E.coli with Cap 2´-O-Methyltransferase gene
Optimum Temperature		37℃
Storage Buffer	20 mM Tris-HCl pH8.0，0.1mM EDTA，1mM DTT，100mM NaCl，50%（v/v）glycerin，0.1%（v/v）Trion X-100
Unit Definition	One unit is defined as the amount of enzyme required to methylate 10 pmol of 80nt capped RNA transcript in 1 h at 37°C.

Contents

Contents No.

Name

Catalog No./Specification

10612ES92

(10 KU)

10612ES97

(50 KU)

10612ES98

(250 KU)

10612ES99

(20 MU)

10612

mRNA Cap 2´-O-Methyltransferase GMP-grade (50 U/μL)

200 μL

1 mL

5 mL

400 mL

Shipping and Storage

The mRNA Cap 2´-O-Methyltransferase GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Experimental methods

Cap1 capping reaction (20 μL reaction system)

This step is suitable for capping reaction of 10 μg RNA (≥ 100 nt), and can be amplified according to experimental needs.

1. Take 10 μg RNA to a 1.5 mL centrifuge tube and dilute to 12.9 μL with nuclease-free water;

2. Heat at 65°C for 5 min;

3. Take out the centrifuge tube and place it on ice for 5 min;

4. Add the following components in sequence:

Components	Volume
Denatured RNA	12.9 μL
10×Capping Buffer	2.0 μL
Murine RNase inhibitor(40 U/μL)	0.5 μL
GTP (10 mM)	1.0 μL
SAM (10 mM, fresh)	1.6 μL
Vaccinia Capping Enzyme (10 U/μL)	1.0 μL
Cap 2´-O-Methyltransferase (50 U/μL)	1.0 μL

【Note】10×Capping Buffer(Cat# 10666)：0.5 M Tris-HCl, 50 mM KCl, 10 mM MgCl₂, 10 mM DTT pH 8.0@25°C.

5. Incubate at 37°C for 2 h;

6. RNA capping is completed, next experiments can be performed.

Notes

1. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

2. The extracted RNA needs to be purified and resuspended in nuclease-free water;

3. The RNA solution needs to be heated before adding the enzyme to remove the secondary structure at the 5'end;

4. For RNA with a known 5'end structure, the reaction time can be extended to 4 h to improve the capping efficiency;

5. In the 5'end labeling reaction system, the GTP stock solution should be diluted to 1-3 times of the mRNA molar concentration in the reaction system.

HB230818

Product Description

This product is produced in accordance with GMP process requirements. The product is provided in liquid form and can be used for Cap1 capping reaction of pre-mRNA in vivo/in vitro.

Product Properties

Source	Recombinant E.coli with Cap 2´-O-Methyltransferase gene
Optimum Temperature		37℃
Storage Buffer	20 mM Tris-HCl pH8.0，0.1mM EDTA，1mM DTT，100mM NaCl，50%（v/v）glycerin，0.1%（v/v）Trion X-100
Unit Definition	One unit is defined as the amount of enzyme required to methylate 10 pmol of 80nt capped RNA transcript in 1 h at 37°C.

Contents

Contents No.

Name

Catalog No./Specification

10612ES92

(10 KU)

10612ES97

(50 KU)

10612ES98

(250 KU)

10612ES99

(20 MU)

10612

mRNA Cap 2´-O-Methyltransferase GMP-grade (50 U/μL)

200 μL

1 mL

5 mL

400 mL

Shipping and Storage

The mRNA Cap 2´-O-Methyltransferase GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Experimental methods

Cap1 capping reaction (20 μL reaction system)

This step is suitable for capping reaction of 10 μg RNA (≥ 100 nt), and can be amplified according to experimental needs.

1. Take 10 μg RNA to a 1.5 mL centrifuge tube and dilute to 12.9 μL with nuclease-free water;

2. Heat at 65°C for 5 min;

3. Take out the centrifuge tube and place it on ice for 5 min;

4. Add the following components in sequence:

Components	Volume
Denatured RNA	12.9 μL
10×Capping Buffer	2.0 μL
Murine RNase inhibitor(40 U/μL)	0.5 μL
GTP (10 mM)	1.0 μL
SAM (10 mM, fresh)	1.6 μL
Vaccinia Capping Enzyme (10 U/μL)	1.0 μL
Cap 2´-O-Methyltransferase (50 U/μL)	1.0 μL

【Note】10×Capping Buffer(Cat# 10666)：0.5 M Tris-HCl, 50 mM KCl, 10 mM MgCl₂, 10 mM DTT pH 8.0@25°C.

5. Incubate at 37°C for 2 h;

6. RNA capping is completed, next experiments can be performed.

Notes

1. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

2. The extracted RNA needs to be purified and resuspended in nuclease-free water;

3. The RNA solution needs to be heated before adding the enzyme to remove the secondary structure at the 5'end;

4. For RNA with a known 5'end structure, the reaction time can be extended to 4 h to improve the capping efficiency;

5. In the 5'end labeling reaction system, the GTP stock solution should be diluted to 1-3 times of the mRNA molar concentration in the reaction system.

HB230818

mRNA Cap 2氧甲基转移酶GMP级|mRNA Cap 2´-O-Methyltransferase GMP-grade

暂无内容

gmp级pei操作方法

发表于2024年9月20日由上海金畔生物科技有限公司

gmp级pei操作方法

Jinpan Polyethyleneimine PEI PRO(GMP)

订购信息

货号

产品名称

规格

78EF10003–5Ml

78EF10003-1L

Jinpan Polyethyleneimine PEI PRO(GMP)

5Ml

储存温度：2-4℃保存⼀年。（避免冷冻）

产品描述

Jinpan Polyethyleneimine PEI PRO(GMP)是GMP级别转染试剂，本制品利BIOHUB公司生产的线性阳离子聚合物,按照符合GMP认证管理体系下，采用药用规格原辅料生产，所用起始材料及所需化学品和配制过程均在无菌环境中操作进行，以确保制造过程中每个步骤的特性、效力、纯度和安全性的一致性。并严格控制重金属残留、细菌内毒素残留等，生产工艺符合 GMP 规范的产品生产与质量管理规程，保障生产过程及所有原辅料可追溯。

操作方法参考

一、贴壁细胞转染以 6 孔板转染 293T 细胞为例：

（一）转染前准备

1) 转染前一天：用胶原酶（WORTHINGTONG 公司 LS004196)消化细胞并计数（不建议用胰酶）。按照每孔2×10 6的细胞量接种293T至6孔板(每孔加入2ml新鲜DMEM:F12+5%FBS *培养基和 1ml 的细胞悬液)置于 37°C,5%CO2培养箱培24h。

2) 24h 后,移除之前培养基，每孔加入 2ml 新鲜的 DMEM:F12+5%FBS。

注意：确保 293T 细胞生长状态良好传代不超过15代。

（二）转染过程

3) 第一天：显微镜下检查细胞汇合度，当细胞达到 70%到 80%的汇合度时，开始准备转染。

4) 对于每孔细胞，用 100µl 无血清培养基（如 OPTI-MEMⅠ培养基）稀释 DNA 使 DNA 终浓度为 1ug/ml（DNA/总培养体积）。

5) 对于每孔细胞，用100μ无血清培养基（如 OPTI-MEMⅠ培养基）稀释适当比例的适量 Jinpan-PRO GMP 。DNA:Jinpan-PRO GMP 的比例要依据自己实验摸索最适比例。

6) 将稀释的 Jinpan-PRO GMP转染试剂加入到稀释的质粒中（总体积 200µL）轻轻混匀，室温孵育30分钟。

7) 小心地将 Jinpan-PRO GMP复合物滴加到细胞培养板每个孔中，轻轻摇动培养板混匀。注意加入混合液时轻轻沿着孔板边缘滴入，而不是在细胞的顶部以免破坏细胞的粘附性。

8) 将培养板放入 37°C，5%CO2培养箱培养中培养 24h。

（三）观察转染结果 9) 第二天：在荧光显微镜下观察细胞转染效率，通常超过 80%的 293T 细胞在转染后的 24h 可以观察到绿色荧光。小贴士 :建议进行不同基因转染时应对 Jinpan-PRO GMP 和 DNA 的比例进行优化，以达到最适比例，通常筛选范围在 1:1 到 1:3 之间。如果之前用过进口品牌的 PEI，用Jinpan-PRO GMP时应适当降低使用浓度。

通常情况下，分别准备Jinpan-PRO GMP 和DNA转染溶液时其体积一般是转染前每孔细胞培养液体积总量的 1/10-1/20，如细胞培养液体积为 3ml，则稀释Jinpan-PRO GMPI及 DNA 的体积为150ul。质粒 DNA 的浓度通常为 1ug/ml（DNA 质量/细胞培养总体积）[1][4]。不同质粒种类以及大小会影响转染效率，如较大片段插入质粒可能转染效率低，可根据实际实验需要调整，如适当增加转染质粒总量等。 ØHEK293 GnTI 细胞重悬浮可用枪头或移液管反复吹打，但 CHO-S 细胞的粘附性比较强重悬浮时需要借助细胞刮刀。一般建议用胶原酶消化细胞，如果表达的是膜蛋白，胰酶消化细胞可能会降低蛋白表达，建议用胶原酶消化细胞，我们推荐(LS004196WORTHINGTONG)的胶原酶。对于贴壁性低的细胞系，可用明胶或鼠尾胶铺板，增加细胞与培养皿之间的粘附性。一旦确定了细胞类型、培养基和78BioPEI GMP与 DNA 的最佳比例，就可以在转染后 24 至 96 小时内多次观察转染效率，以优化最大表达量时间点。

二、悬浮细胞转染离心对数期生长的细胞用新鲜的培养基重悬浮使细胞密度达到 1×10 6cells/mL 小规模转染时，如需大规模转染细胞密度要到达 2-3×10 6cellsml/mL，可参考文献[4]。以 293F 细胞于 100mL 培养瓶（溶液体积占瓶体积的 1/5）操作体系举例如下：

（一）转染前准备

1) HEK293F 细胞传代接种于 20mL 悬浮细胞生长培养基中(36.5℃，120rpm，5%CO2 )的条件下培养。当细胞密度到 1X10 6cells/mL，然后旋紧瓶口放入摇床继续培养，2~4 小时后可以进行转染。

（二）转染过程

2) 用 1mlOptiPRO™SFM(无血清培养基)稀释适量的 DNA，使 DNA 终浓度为 1ug/ml（DNA/总培养体积）。混匀并静置 5min。

3) 用1mlOptiPRO™SFM(无血清培养基)稀释适当比例的Jinpan-PRO GMP，混匀并静置 10min。

（78BioPEI：DNA 参考比例范围，可参考上述贴壁细胞Jinpan-PRO GMP和 DNA 优化方案优化）。

4) 将Jinpan-PRO GMP转染试剂加入到稀释的质粒中轻轻混匀，室温孵育 30 分钟。

5) 将Jinpan-PRO GMP转染复合物逐滴加入到细胞培养液中，摇匀后旋紧瓶口放回摇床（36.5℃，5%CO2,120rpm）。

6) 基因表达可在 24-72 小时后检测，具体时间取决于细胞系和转基因.

小贴士

在悬浮培养中，单个细胞的转染效率要高于聚集在一起的细胞，需要优化适合单细胞的生长条件。方形瓶应经过两个连续的干燥循环高压灭菌（每个 45 分钟，干燥 15 分钟）盖子应尽可能松，不得脱落。应该让它们冷却拧紧盖子前，将其*放入层流罩中。如果瓶子向内塌陷，细胞将无法正常生长。如果要获得更多的蛋白产量，有参考文献表明转染后 24 小时，可以适当稀释细胞，稀释比例参考范围（1:2—1:5）之间，可以增加蛋白产量，稀释效应与最佳稀释比率应根据经验确定, 可以把 DNA 和Jinpan-PRO GMP 转染试剂直接加入到细胞悬液中进行转染，但必须经过上述特定流程的 DNA 和转染试剂的比例和用量的相应优化。

10×转录缓冲液(GMP级,不含Mg2+)

发表于2024年7月2日由上海金畔生物科技有限公司

10×转录缓冲液(GMP级,不含Mg2+)

产品说明书

FAQ

COA

已发表文献

This product is optimized transcription buffer and efficient for in vitro synthesis of RNA. The formulation of the product: 400mM Tris-HCl, 20mM spermidine, 100mM DTT, pH 7.9 at 25℃.

This product is produced in accordance with GMP regulations and provided in liquid form.

Contents

Contents No.	Name	Catalog No./Specification
Contents No.	Name	10669ES03	10669ES10	10669ES60
10669	10×Transcription Buffer GMP-grade (Mg²⁺ free)	1 mL	10 mL	100 mL

Shipping and Storage

This product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB220701

This product is optimized transcription buffer and efficient for in vitro synthesis of RNA. The formulation of the product: 400mM Tris-HCl, 20mM spermidine, 100mM DTT, pH 7.9 at 25℃.

This product is produced in accordance with GMP regulations and provided in liquid form.

Contents

Contents No.	Name	Catalog No./Specification
Contents No.	Name	10669ES03	10669ES10	10669ES60
10669	10×Transcription Buffer GMP-grade (Mg²⁺ free)	1 mL	10 mL	100 mL

Shipping and Storage

This product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB220701

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无机焦磷酸酶Pyrophosphatase GMP级(0.1 U/μL)

发表于2024年6月23日由上海金畔生物科技有限公司

无机焦磷酸酶Pyrophosphatase GMP级(0.1 U/μL)

产品说明书

FAQ

COA

已发表文献

This product is an inorganic pyrophosphatase derived from recombinant expression in E. coli. It is an enzyme that can catalyze the conversion of one molecule of pyrophosphate into two molecules of phosphate ions. This is a high-energy reaction, so this reaction can be coupled to some thermodynamically unfavorable transformations in order to drive these transformations to completion. Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to orthophosphate. In molecular biology, it can be used to increase RNA yield in reverse transcription reaction.

This product is produced in accordance with GMP process requirements, and the product is provided in liquid form

Product Properties

Source	Recombinant E.coli with Pyrophosphatase gene from yeast
Optimum Temperature	25℃
Storage Buffer	50 mM Tris-HCl pH 7.9, 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% （v/v）TritonX-100, 50%（v/v）glycerin
Unit Definition	One unit (U) is the amount of enzyme needed to catalyze the hydrolysis of PPI per minute to produce 1 μmol Pi.（The standard reaction is: 500 μL system contains 100 mM Tris-HCl pH7.2, 2 mM MgCl₂, 2 mM Ppi; reaction at 25℃ for 10 min）

Contents

Catalog No.

Name

Catalog No./Specification

10672ES03

(1 U)

10672ES10

(10 U)

10672ES60

(100 U)

10672

Pyrophosphatase, Inorganic GMP-grade (0.1 U/μL)

10 μL

100 μL

1 mL

Shipping and Storage

The Pyrophosphatase, Inorganic GMP-grade products shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. This product is active in a variety of reaction buffers. Usually, this product can be directly connected in experiments such as IVT.

2.The optimal reaction temperature of this product is 25℃, it is active at 16-37℃, 65℃ can inactivate the enzyme for 10 minutes.

3. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this

HB220701

This product is produced in accordance with GMP process requirements, and the product is provided in liquid form

Product Properties

Source	Recombinant E.coli with Pyrophosphatase gene from yeast
Optimum Temperature	25℃
Storage Buffer	50 mM Tris-HCl pH 7.9, 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% （v/v）TritonX-100, 50%（v/v）glycerin
Unit Definition	One unit (U) is the amount of enzyme needed to catalyze the hydrolysis of PPI per minute to produce 1 μmol Pi.（The standard reaction is: 500 μL system contains 100 mM Tris-HCl pH7.2, 2 mM MgCl₂, 2 mM Ppi; reaction at 25℃ for 10 min）

Contents

Catalog No.

Name

Catalog No./Specification

10672ES03

(1 U)

10672ES10

(10 U)

10672ES60

(100 U)

10672

Pyrophosphatase, Inorganic GMP-grade (0.1 U/μL)

10 μL

100 μL

1 mL

Shipping and Storage

The Pyrophosphatase, Inorganic GMP-grade products shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Notes

1. This product is active in a variety of reaction buffers. Usually, this product can be directly connected in experiments such as IVT.

2.The optimal reaction temperature of this product is 25℃, it is active at 16-37℃, 65℃ can inactivate the enzyme for 10 minutes.

3. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this

HB220701

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GTP Tris溶液GMP级(100 mM) GTP Tris Solution

发表于2024年6月22日由上海金畔生物科技有限公司

GTP Tris溶液GMP级(100 mM) GTP Tris Solution

产品说明书

FAQ

COA

已发表文献

Product Information

Product name	Cat#	Size
GTP Tris Solution GMP-grade (100 mM)	10655ES03	1 mL
	10655ES10	10 mL
	10655ES60	100 mL

Product Description

GTP, Guanosine-5'-triphosphate, can be used in various of molecular biology applications, such as in vitro transcription, RNA amplification, siRNA synthesis, etc. In addition, GTP plays an important role in signal transduction as a phosphate or pyrophosphate messenger: GTP can activate G proteins, which can induce multiple protein kinase mediated cascade reactions, causing a variety of cytological behaviors, such as cell proliferation, differentiation, etc. GTP can also be used as a high-energy precursor of single nucleotides to participate in the synthesis of DNase and RNase.

This product is a transparent colorless aqueous solution prepared with GTP tris solution, and free of DNase and RNase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	86-01-1 (free acid)
Formula	C₁₀H₁₆N₅O₁₄P₃(free acid)
Molecular weight	523.18 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

Product Information

Product name	Cat#	Size
GTP Tris Solution GMP-grade (100 mM)	10655ES03	1 mL
	10655ES10	10 mL
	10655ES60	100 mL

Product Description

This product is a transparent colorless aqueous solution prepared with GTP tris solution, and free of DNase and RNase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	86-01-1 (free acid)
Formula	C₁₀H₁₆N₅O₁₄P₃(free acid)
Molecular weight	523.18 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

GTP Tris溶液GMP级(100 mM) GTP Tris Solution

暂无内容

UTP Tris溶液GMP级(100 mM) UTP Tris Solution

发表于2024年6月21日由上海金畔生物科技有限公司

UTP Tris溶液GMP级(100 mM) UTP Tris Solution

产品说明书

FAQ

COA

已发表文献

Product Information

Product name	Cat#	Size
UTP Tris Solution GMP-grade (100 mM)	10654ES03	1 mL
	10654ES10	10 mL
	10654ES60	100 mL

Product Description

UTP, Uridine-5'-triphosphate, can be used in a variety of molecular biology applications, such as in vitro transcription, RNA amplification, siRNA synthesis, etc. In addition, UTP is involved in galactose metabolism. The activated form of UDP-galactose is converted to UDP-glucose, which participates in the glycogen synthesis pathway.

This product is a transparent colorless aqueous solution prepared from UTP tris solution, and free from DNase and RNase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	63-39-8 (free acid)
Formula	C₉H₁₅N₂O₁₅P₃(free acid)
Molecular weight	484.14 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

Product Information

Product name	Cat#	Size
UTP Tris Solution GMP-grade (100 mM)	10654ES03	1 mL
	10654ES10	10 mL
	10654ES60	100 mL

Product Description

This product is a transparent colorless aqueous solution prepared from UTP tris solution, and free from DNase and RNase contamination.

This product is produced in accordance with GMP process requirements and provided in liquid form.

Product Properties

CAS No	63-39-8 (free acid)
Formula	C₉H₁₅N₂O₁₅P₃(free acid)
Molecular weight	484.14 g/moL (free acid)
Purity(HPLC)	≥ 99%
Content	100 mM ± 3 mM
Structure

Shipping and Storage

The product is shipped with dry ice and can be stored at -25 ~ -15℃ for two years.

Cautions

Keep on ice while using and return immediately to -20°C after use.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB240125

UTP Tris溶液GMP级(100 mM) UTP Tris Solution

暂无内容