首页 > 蛋白纯化和免疫沉淀的磁珠系列 > Protein A/G免疫沉淀磁珠

Protein A/G免疫沉淀磁珠

产品号

规格

单价

数量

B23201	1 mL	RMB 438.76
B23202	5 mL	RMB 1800.08

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产品优势
实验数据
价格对比
产品描述
储存条件
实验方法
资料下载

产品优势

省时：比亲和凝胶法节省大约80min（以IP实验为例比较）；
操作简单：磁性分离，无需离心；
非特异低：亲和凝胶孔径大，容易产生非特异吸附，而磁珠孔径小，不容易产生非特异吸附；
结合量高：磁珠表面PorteinA/G的蛋白包被量高达9.3×1013个分子/平方厘米，抗体加载量为0.6-0.8 mg/mL磁珠悬液。

实验数据

Customer Product Validation(10)

Septin4 is a novel PARP1 interacting protein and the interaction is enhanced under oxidative stress. (A) Immunoprecipitation (IP) and western blotting (IP-western) using anti-PARP1 antibody or negative control IgG and Protein A/G immunoprecipitation magnetic beads followed by anti-Septin4 western blot were performed to ascertain endogenous interaction between PARP1 and Septin4. (B) Anti-Septin4 antibody or negative control IgG and Protein A/G immunoprecipitation magnetic beads followed by anti-PARP1 western blot were performed to ascertain endogenous interaction between Septin4 and PARP1. (C) Endogenous interaction between PARP1 and Septin4 was enhanced by treatment of H2O2, which were evaluated by Immunoprecipitation using anti-PARP1 antibody. (D) Endogenous interaction between Septin4 and PARP1 was enhanced by treatment of H2O2, which were evaluated by Immunoprecipitation using anti-Septin4 antibody.
Exogenous HA-Flag-PHF5A in HEK 293T cells was immunoprecipitated with anti-Flag or control IgG. The immunoprecipitates were then analyzed by Western blot.
The ubiquitination level of Mfn1, Mfn2 and Cx43 in cardiomyocytes which were transfected with gp78-siRNA or Con-siRNA and treated by 5 μM NPS R568 or 3 μM Calhex231 respectively by immunoprecipitation (n= 3). A representative blot is shown. *P<0.05 versus control; # P<0.05 versus HG
(F) Whole-cell lysates from osteosarcoma cell lines were immunoprecipitated with an anti-CSE1L antibody followed by immunoblotting (IB) with anti-MSH6 and anti-CSE1L antibodies. IgG was used as a negative control. (G) Whole-cell lysates from osteosarcoma cell lines were immunoprecipitated with an anti-MSH6 antibody followed by IB with anti-CSE1L and anti-MSH6 antibodies. IgG was used as a negative control.
Immunoprecipitation for c-Myc protein:
100 μg of Protein A/G Magnetic beads & 10 μg anti-c-Myc antibody was used.
Immunoprecipitation for Flag Tag-fusion Protein:
1. 1% input
2. Biotool Magnetic Protein A/G beads
3. Biotool Magnetic Protein A/G beads + Mouse Anti-Flag Antibody
4. Life technology Magnetic Protein A/G beads + Mouse Anti-Flag Antibody + IP
5. Biotool Magnetic Protein A/G beads + Mouse Anti-Flag Antibody + IP
6. Santa cruz Protein A/G Agarose gel + Mouse Anti-Flag Antibody + IP
Immunoprecipitation for HAS:
1. 5 ug of rabbit polyclonal antibody was used
2. 5 ug of mouse IgG1 monoclonal antibody was used
Immunoprecipitation for GST-Tag Fusion Protein:
100 ug of Biotool Protein A/G Magnetic beads & 10 ug Anti- GST-Tag Antibody was used
(I and J) 293T cells were co-transfected with a HA-tagged TGEV S1 expression plasmid together with GFP-tagged EGFR Receptor 1 or GFP-tagged EGFR Receptor 2 expression plasmid, cell lysates were immunoprecipitated with an anti-HA antibody or an anti-GFP antibody, the resulting precipitates were examined by immunoblotting using an anti-HA or an anti-GFP antibody to examine the interaction between HA-TGEV S1 and GFP-tagged EGFR. (** p < 0.01).
Immunoprecipitation for HAS:
100 ug of Biotool Protein A/G Magnetic beads & 10 ug Anti-HAS Antibody was used
Immunoprecipitation for GST-Tag Fusion Protein:
1. Antigen & Anti-GST-Tag Antibody (Mouse IgG1)
2 & 3. Elution after IP
4. Antigen & Anti-GST-Tag Antibody (Mouse IgG1)
5 & 6. Elution after IP (Control group without GST-Tag Fusion Protein)

价格对比

产品描述

Protein A/G免疫沉淀磁珠使用“纳米表面生物技术”(S-TEC)，Protein A/G包被在超顺磁性磁珠微球表面，包被密度高达9.3×10¹³个分子/平方厘米。与同类产品相比，Bimake Protein A/G免疫沉淀磁珠表面具有更多的抗体结合位点。在IP实验中，只需使用更少的磁珠量，非特异性结合低，使用简便有效。Bimake Protein A/G免疫沉淀磁珠具有大面积特异的表面区域，从而大大缩短抗体吸附和抗原结合的时间。最短时，抗原和抗体的结合时间只需10分钟，可以在30分钟内完成完整的IP实验。Bimake Protein A/G免疫沉淀磁珠适用的范围广泛，例如细胞裂解液、细胞分泌液上清、血清、动物腹水以及其它的免疫抗原等。

特性

Protein A/G是融合了Protein A和Protein G蛋白的IgG结合区的重组融合蛋白，它包含了Protein A的四个Fc结合区域以及Protein G的两个Fc结合区域，分子量50460 Da。相比Protein A，Protein A/G的结合更少依赖pH值，但是却具有了Protein A和G两者的加合效应。

产品性质

浓度	10 mg/mL
磁珠大小	100 nm
人IgG结合能力	0.4-0.5 mg/mL
配体	重组蛋白A/G
应用	rProtein纯化，免疫沉淀
pH稳定性	6-8 （长期）
运输条件	蓝冰

储存条件

4°C可以储存2年。在缺乏甘油的情况下避免反复冻融。

实验方法

图1展示了免疫共沉淀（IP）的操作方法。可以根据抗体类型和靶蛋白进行进一步的优化。Protein A/G免疫磁珠是进行IP实验的最佳方案。

Figure 1 General Protocol for Immunoprecipitation

表1 免疫共沉淀（IP）的操作方法

表2. 推荐的缓冲液配方
缓冲液	推荐配方（自备）
Binding buffer	50 mM Tris, 150 mM NaCl, 0.1%-0.5% detergent (TritonX-100, Tween 20 or NP40), pH 7.5
Wash buffer	50 mM Tris, 150 mM NaCl, 0.1%-0.5% detergent, pH 7.5
Elution buffer	0.1 M -0.2 M Glycine, 0.1%-0.5% detergent, pH 2.5-3.1 (or 0.1 M citric acid, 0.1%-0.5% detergent, pH 2.5-3.1)
Neutralize buffer	1 M Tris, pH8.0

实验之前的注意事项

1 进行IP实验之前，请仔细阅读操作说明；
2 该产品需要使用磁力架
3 使用之前，请将Protein A/G免疫磁珠混匀；
4 Protein A/G免疫磁珠应该保持在储存液中避免干掉；
5 Protein A/G免疫磁珠应避免冻融或离心；
6 为保证最佳结果，请选择特异性好的抗体；
7 根据不同的抗体和抗原仅进行实验方法的优化；

表1. 抗体与Protein A/G及Protein A的结合能力比较
Species	Antibody class	sProtein A/G	sProtein A
Human	Total IgG	＋＋＋＋＋	＋＋＋＋＋
	IgG1, IgG2	＋＋＋＋＋	＋＋＋＋＋
	IgG3	＋＋＋＋＋	＋
	IgG4	＋＋＋＋＋	＋＋＋＋＋
	IgM	＋	＋
	IgD	－	－
	IgA	＋	＋
	IgA1, IgA2	＋	＋
	IgE	＋＋＋	＋＋＋
	Fab	＋	＋
	ScFv	＋	＋
Mouse	Total IgG	＋＋＋＋＋	＋＋＋＋＋
	IgM	－	－
	IgG1	＋＋＋	＋
	IgG2a	＋＋＋	＋＋＋
	IgG2b	＋＋＋	＋＋＋
	IgG3	＋＋＋	＋＋＋＋＋
Rat	Total IgG	＋＋＋	＋
	IgG 1	＋＋＋	＋＋＋
	IgG2a	＋＋＋＋＋	＋＋＋
	IgG2b	＋	＋＋＋
	IgG2c	＋＋＋＋＋	＋＋＋
Cow	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Goat	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Sheep	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Horse	Total IgG	＋＋＋＋＋	＋
	IgG(ab), IgG(c)	＋	－
	IgG(T)	＋＋＋＋＋	＋
Rabbit	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Guinea pig	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Hamster	Total IgG	＋＋＋	＋＋＋
Pig	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Donkey	Total IgG	＋＋＋＋＋	＋＋＋
Cat	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Dog	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Monkey	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Chicken	Total IgG	－	－