Anti-Hsp27 (phospho S78) antibody [Y175] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 (phospho S78) antibody [Y175] - BSA and Azide free (AB247262)

This data was developed using ab32501, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling Hsp27 with purified ab32501 at 1/5000 dilution (0.11 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 (phospho S78) antibody [Y175] - BSA and Azide free (AB247262)

This data was developed using ab32501, the same antibody clone in a different buffer formulation.

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25 ug/mL anisomycin for 30 min, then Lambda Protein Phosphatase 31 for 2 hours cells labeling Hsp27 with purified ab32501 at 1/50 dilution (11.3 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/mL) dilution. Goat anti-rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-Hsp27 (phospho S78) antibody [Y175] - BSA and Azide free (AB247262)

This data was developed using ab32501, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Hsp27 (phospho S78) antibody [Y175] (<a href='/en-us/products/primary-antibodies/hsp27-phospho-s78-antibody-y175-ab32501'>ab32501</a>) at 1/5000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 250 ng/ml anisomycin for 30 minutes whole cell lysate at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 250 ng/ml anisomycin for 30 minutes whole cell lysate, and then the menbrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 27 kDa

false

• Dot

Unknown

Dot Blot - Anti-Hsp27 (phospho S78) antibody [Y175] - BSA and Azide free (AB247262)

This data was developed using ab32501, the same antibody clone in a different buffer formulation.Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using ab32501 at dilution 1/2000.