PA5-17662

antibody from Invitrogen Antibodies

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

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-17662

Provider

Invitrogen Antibodies

Product name

Phospho-HSP27 (Ser78) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho Heat Shock Protein 27 (p-HSP27) pSer78 (green) in HeLa cells either left untreated (left panel) or treated with 25 µg/mL anisomycin for 30 min (right panel). 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 (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Phospho-HSP27 pSer78 polyclonal antibody (Product # PA5-17662) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry was performed on paraffin-embedded human breast carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 10 minutes at 95C. Following antigen-retrieval, tissues were incubated in 3% hydrogen peroxide for 10 minutes, washed, and then blocked in 5% normal goat serum in TBST for 1 hour at room temperature. Tissues were stained with a phospho-HSP27 pSer78 polyclonal antibody (Product # PA5-17662) at a dilution of 1:100 overnight at 4C. Tissues were washed extensively with TBS + 0.1% Tween-20 and detection was performed using an HRP-conjugated secondary reagent followed by DAB substrate.