Anti-SH2-B alpha / beta / gamma / delta Antibody (46) recommended for detection of SH2-B α, β, γ and δ of mouse, rat and human origin by WB, IP and IF

Anti-SH2-B α/β/γ/δ Antibody (46): sc-136065

Anti-SH2-B α/β/γ/δ Antibody (46) is rated 5.0 out of 5 by 3.
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Datasheets

    • SH2-B α/β/γ/δ Antibody (46) is a mouse monoclonal IgG1, cited in 4 publications, provided at 50 µg/0.5 ml
    • raised against amino acids 403-513 of SH2-Bβ of rat origin
    • recommended for detection of SH2-B α, β, γ and δ of mouse, rat and human origin by WB, IP and IF
    • At present, we have not yet completed the identification of the preferred secondary detection reagent(s) for SH2-B α/β/γ/δ Antibody (46). This work is in progress.
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I am using SH2-B α/β/γ/δ (46): sc-136065 for immunofluorescence, what dilution of this antibody should I use?

Asked by: DefinitelyNotMatt
Thank you for your question. We recommend a starting dilution of 1:50. However, optimal antibody concentration should be determined by titration. Please find our full immunofluorescence protocol here: https://www.scbt.com/scbt/resources/protocols/immunofluorescence-cell-staining
Answered by: Technical Support
Date published: 2017-03-24
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Rated 5 out of 5 by from Immunoblots endogenous SH2B1 in PC12 cells Anti-SH2B1 (46) against aa 403-513 of rat SH2B1 (sc. 136065) PC12 cells stably expressing GFP tag (only endogenous SH2B1 present). Treated ± NGF (100 ng/ml), cells lysed in RIPA buffer (1% Triton X100). Load 40 ul on minigel. Immunoblot: block with 3% BSA in TBS-1% tween, blot with anti-SH2B1 (46) (1/250 dilution). 2o antibody: mIGGkBP-HRP (sc) (1/1000). Nice SH2B1 signal detected with NGF-dependent shift in migration. Band at 90 kDa in absence of NGF. Band at 100 kDa in sample treated with NGF. 293T cell transfected with the four isoforms of human GFP-SH2B1. Cells lysed in RIPA buffer (1% Triton X100). Load 20 ul on minigel. Immunoblot: block with 3% BSA in TBS-1% tween, blot with anti-SH2B1 (46) (1/1000 dilution). 2o antibody: anti-mouse IRdye680 (1/20,000). The four isoforms of SH2B1 were detected by the SH2B1 (46) antibody. For most antibodies prepared against SH2B1, the background is diminished when a 2o antibody conjugated to HRP is used (when compared to 2o antibodies conjugated to IRdyes.)
Date published: 2018-09-12
Rated 5 out of 5 by from Produced excellent immunofluorescence nuclear Produced excellent immunofluorescence nuclear and cytoplasmic staining in HeLa cells. -SCBT QC
Date published: 2015-03-23
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