Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using ab62339, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human skin labeling Hsp27 with ab62339 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Cytoplasmic staining on human skin.
The section was incubated with ab62339 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using ab62339, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human breast cancer labeling Hsp27 with ab62339 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Cytoplasmic staining on human breast cancer.
The section was incubated with ab62339 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using ab62339, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat skin labeling Hsp27 with ab62339 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Cytoplasmic staining on rat skin.
The section was incubated with ab62339 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using ab62339, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse skin labeling Hsp27 with ab62339 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Cytoplasmic staining on mouse skin.
The section was incubated with ab62339 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Lab

Western blot - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using the same antibody clone in a different buffer formulation (ab62339).

Lanes 1- 2 : Merged signal (red and green). Green - ab62339 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab62339 was shown to react with Hsp27 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261738 (knockout cell lysate ab256945) was used. Wild-type HeLa and HSPB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab62339 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hsp27 antibody [EP1724Y] (<a href='/en-us/products/primary-antibodies/hsp27-antibody-ep1724y-ab62339'>ab62339</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HSPB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HSPB1 (Hsp27) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-hspb1-hsp27-knockout-hela-cell-line-ab261738'>ab261738</a>)

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

• WB

Lab

Western blot - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (AB247352)

This data was developed using the same antibody clone in a different buffer formulation (ab62339).

Lanes 1 - 4 : Merged signal (red and green). Green - ab62339 observed at 27 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab62339 was shown to specifically react with Hsp27 in wild-type HAP1 cells. No band was observed when Hsp27 knockout samples were used. Wild-type and Hsp27 knockout samples were subjected to SDS-PAGE. ab62339 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hsp27 antibody [EP1724Y] - BSA and Azide free (ab247352) at 1/500 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

Hsp27 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

MCF7 whole cell lysate at 20 µg

Predicted band size: 23 kDa

false