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Immunoprecipitation Kit (Protein A+G Agarose Gel)

 
Catalog No.
K4623
经典Protein A+G琼脂糖IP试剂盒,用于免疫(共)沉淀、蛋白分子互作等方面
组合的产品项目
规格价格库存 数量
20T
¥ 1,406.00
现货

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B

Description

免疫沉淀(Immunoprecipitation,IP)或免疫共沉淀(Co-Immunoprecipitation,Co-IP)是研究蛋白或蛋白与蛋白间相互作用(Protein-Protein Interactions, PPIs)的常用技术手段,通过使用特异性抗体和可结合抗体的介质(如Protein A/G琼脂糖凝胶等),或直接使用偶联特异性抗体的介质(如琼脂糖凝胶或磁珠),将目标蛋白质从复杂样品中分离出来,后续可用于Western印迹检测或质谱分析等。

Protein A是一种发现于金黄色葡萄球菌(Staphylococcus aureus)的细胞壁表面蛋白,分子量为42 kDa,能特异性地与哺乳动物免疫球蛋白(Immunoglobulin, Ig) Fc区结合,同时也会和人VH3家族的Fab区结合。Protein G是C型或G型链球菌(Streptococcal bacteria)表达的免疫球蛋白结合蛋白,能特异性地与哺乳动物免疫球蛋白(Immunoglobulin, Ig)Fc区结合。Protein A可特异性地结合相应抗体,如human IgG1、IgG2、IgG4,mouse IgG2a、IgG2b及rabbit IgG等,且每个Protein A分子可以结合5个IgG分子,而Protein G可结合的抗体为human IgG1、IgG2、IgG3、IgG4,mouse IgG1、IgG2a、IgG2b、IgG3、rat IgG1、IgG2a、IgG2b、IgG2c,以及rabbit、goat多克隆抗体等,且每个Protein G分子可以结合3个IgG分子。针对常见的免疫球蛋白亚类的结合能力及不同物种的总结合能力情况见下表(Table 1)。

本试剂盒包含Protein A+G琼脂糖凝胶及其他免疫沉淀必要试剂,配合用户自备的特异性抗体,使免疫沉淀(Immunoprecipitation, IP,也称Pull-down)或免疫共沉淀(Co-IP)实验更加简单、便捷、高效,免疫沉淀产物可用于目的蛋白或其蛋白复合物的检测等实验。(具体产品参数见Table 2)

Species

Subclass

Protein A binding

Protein G binding

Human

lgA

++

-

lgD

++

-

IgE

++

-

lgG1

++++

++++

lgG2

++++

++++

IgG3

-

++++

lgG4

++++

++++

lgM

++

-

Mouse

IgG1

+

++++

lgG2a

++++

++++

lgG2b

+++

+++

lgG3

++

+++

lgM

+/-

-

Rat

lgG1

-

+

lgG2a

-

++++

lgG2b

-

++

lgG3

+

++

Avian egg yolk

IgY

-

-

Cow

 

++

++++

Dog

 

++

+

Goat

 

-

++

Guinea Pig

lgG1

++++

++

lgG2

++++

++

Hamster

 

+

++

Horse

 

++

++++

Koala

 

-

+

Llama

 

-

+

Monkey (rhesus)

 

++++

++++

Pig

 

+++

+++

Rabbit

 

++++

+++

Sheep

 

+/-

++

Table 1 affinity data for Protein A and Protein G for different sources and subtypes of IgG. ++++ = Strong Binding; ++~+++ = Medium Binding; + = Weak Binding; +/- = Weak or No Binding; - = No Binding.

Characteristics

Description

Product content

Store in 20% ethanol with a glue-to-suspension ratio of approximately 75%

Bead structure

4% cross-linked agarose

Average beads size

90 μm

Coupled protein

Recombinant Protein A and Protein G

M. W. of protein

14 kDa Protein A and 22 kDa Protein G

Ligand concentration

6 mg rProtein A and 2 mg rProtein G per mL settled gel

Binding capacity

≥25 mg human IgG per ml settled gel

Elution method

Elution with acid, competing peptide or SDS-PAGE loading buffer

Application

Suitable for IP, Co-IP, Protein purification

Storage

4°C

Table 2 The main related indicators of Protein A+G Agarose Gel.

质量控制

质量控制和DataSheet

批次:

相关生物数据

Immunoprecipitation Kit (Protein A+G Agarose Gel)
 

Components and Storage

Components K4623-20 T
Cell Lysis Buffer 50 mL
Protease Inhibitor Cocktail (EDTA-Free,100X in DMSO) 0.5 mL
10X TBS 10 mL
Neutralization Buffer 200 μL
Acid Elution Buffer 2 mL
Protein A+G Agarose Gel 400 μL
5X Protein Loading Buffer (Reducing) 600 μL

Store Protease Inhibitor Cocktail (EDTA-Free,100X in DMSO) and 5X Protein Loading Buffer (Reducing) at -20°C and the rest of components at 4°C for 12 months.