Beads抗体-Anti-HA-Tag Mouse Antibody(Agarose Conjugated)

Datasheet

Description

Anti-HA-Tag Mouse mAb (Agarose Conjugated) is a monoclonal anti-HA

antibody covalently linked to agarose; the agarose enables immunoprecipitation (IP) of HA tagged proteins or co-immunoprecipitation (Co- IP) of their interacting partners.


Source

This Abmart monoclonal antibody is produced by immunizing animals with a synthetic peptide containing the influenza hemagglutinin epitope (YPYDVPDYA) (KLH-coupled).


Specificity

Anti-HA-Tag Mouse mAb detects transfected proteins containing the HA epitope tag.



Storage

The product is supplied as a 50% slurry in storage buffer (1 PBS, pH 7.4, containing 0.1% NaN ). Store the product at 4°C and do not freeze.


Reactivity

All

Isotype

Recommended elution buffer

0.2 M Glycine, pH 2.2

Immunoprecipitation procedure

The work can be performed in 1.5 ml micro-centrifuge tubes or in spin columns.

1. Thoroughly resuspend the Anti-HA Agarose by inverting the tube or vial several times.

2. Add 20-50 μl 50% slurry of Anti-HA Agarose into cell lysate using a widebore pipette tip.

Note: The lysate should be fresh, and for a well expressed tagged protein, 200 μl lysate (200-500 μg total protein) usually yields a good IP result.

3. Incubate with gentle mixing for 2 h to overnight at 4 .

4. Wash the beads with 1 ml TBS buffer or lysis buffer, such as RIPA (50 mM Tris HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate), centrifuge for 3 min at 2,000 g, and discard the supernatant. Wash 3 times, avoid losing beads during washes.

5. Elution of the HA tagged protein.

Option 1. Elution with elution buffer.

Add 30-50 μl elution buffer to the beads, gently tap the tube to mix well, immediately centrifuge for 3 min, transfer the supernatant very carefully to a fresh tube (Avoid transferring any beads).

Note: Neutralize the eluant immediately by add 1 l of 1.5 M Tris, pH 9.0 per 20 μl Elution buffer.

Option 2. Elution with HA peptide

Add 30-50 μl HA peptide solution (100 μg/ml HA peptide in TBS buffer), gently tap the tube to mix well, incubate for 10 min, centrifuge for 3 min, and transfer the supernant to a fresh tube. TBS buffer: 50 mM Tris HCl, 150 mM NaCl, pH 7.4.

Option 3. Elution with SDS loading buffer

Add 30 μl 2 SDS loading buffer, gently tap the tube to mix well, boil at 100 °C for 5 min, centrifuge for 3 min, transfer the supernatant to a fresh tube.

Note: in this case, the supernatant contains not only the binding proteins, but also IgG (heavy and light chains).

6. Prepare SDS-PAGE gel for western blotting or proceed to other assays.

Companion products

#M20001 His-Tag (2A8) Mouse mAb

#M20002 Myc-Tag (19C2) Mouse mAb

#M20003 HA-Tag (26D11) Mouse mAb

#M20004 GFP-Tag (7G9) Mouse mAb

#M20007 GST-Tag (12G8) Mouse mAb

#M20008 DYDDDDDK-Tag (3B9) Mouse mAb (Binds to same epitope as

Sigma’s Anti-FLAG M2 Antibody)

#M20012 Anti-Myc-Tag Mouse mAb (Agarose Conjugated)

#M20018 Anti-DYKDDDDK-Tag Mouse mAb (Agarose Conjugated) (Binds to

same epitope as Sigma’s Anti-FLAG M2 Antibody)


M20013

货号 规格 价格(¥)
订购咨询 orders@ab-mart.com
技术支持 service3@ab-mart.com

订货热线 4006-123-828

经销商名录:点击查看


banner-500.jpg

艾比玛特医药科技(上海)有限公司
上海市徐汇区桂平路333号聚科生物园区1号楼1-3层

邮箱:market@ab-mart.com

应聘职位:hr@ab-mart.com

订购专线:4006-123-828

销售电话:13162127138

技术支持:13162477137

总机:021-34695901


wx.png

      微信客服

邮箱:market@ab-mart.com

总机:021-34695901

热线:13166434152 

订购专线:4006-123-828

wx.png

微信客服

沪ICP备17056956号-2    艾比玛特医药科技(上海)有限公司