GTX25581

antibody from GeneTex

Targeting: HSPB1 CMT2F, Hs.76067, Hsp25, HSP27, HSP28

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Data presented on provider website)

Antibody Data

Product number

GTX25581

Provider

GeneTex

Proper citation

GeneTex Cat#GTX25581, RRID:AB_371723

Product name

HSP27 (phospho Ser15) antibody

Antibody type

Polyclonal

Reactivity

Human, Rat, Rabbit

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Western blot analysis of Phospho Heat Shock Protein 27 (p-HSP27) pSer15 was performed by loading 50ug of HeLa cell lysates from untreated cells (left lane) or cells treated with 10uM Anisomycin (right lane) for 30 minutes and 15ul onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a p-HSP27 pSer15 polyclonal antibody at a dilution of 1:500 overnight at 4¢XC on a rocking platform, washed with TBS-0.1%Tween-20, and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed.

Immunocytochemistry

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho Heat Shock Protein 27 (p-HSP27) pSer15 (green) in HeLa cells either left untreated (left panel) or treated with 10uM Anisomysin (right panel) for 30 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a p-HSP27 pSer15 polyclonal antibody at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin and nuclei (blue) were stained with Hoechst dye.

Immunoprecipitation

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Immunoprecipitation of Phospho Heat Shock Protein 27 (p-HSP27) pSer15 was performed on HeLa cells treated with 10uM Anisomysin for 30 minutes. Antigen-antibody complexes were formed by incubating 500ug of whole cell lysate with 3ug of p-HSP27 pSer15 polyclonal antibody (GTX25581) overnight on a rocking platform at 4¢XC. The immune complexes were captured on 50ul Protein A/G Agarose, washed extensively, and eluted. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a p-HSP27 pSer15 polyclonal antibody (GTX25581) at a dilution of 1:500 overnight rotating at 4¢XC, washed with TBST, and probed with an IP detection reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed