Anti-Bak antibody [7D10] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (AB252251)

This data was developed using ab243151 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Bak with ab243151 at 1/50 dilution followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Cytoplasmic staining on human breast carcinoma. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (AB252251)

This data was developed using ab243151 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling Bak with ab243151 at 1/50 dilution followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Cytoplasmic staining on human colon carcinoma. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (AB252251)

This data was developed using ab243151, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12881569).

This antibody reacts with an unidentifiable protein around 150kDa.

Exposure time : 1 second.

All lanes:

Western blot - Anti-Bak antibody [7D10] (<a href='/en-us/products/primary-antibodies/bak-antibody-7d10-ab243151'>ab243151</a>) at 1/5000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

T47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate at 10 µg

Lane 6:

Human heart tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 23 kDa

Observed band size: 20 kDa,25 kDa

false

• WB

Lab

Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (AB252251)

This data was developed using ab243151, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1-5 : Merged signal (red and green). Green - ab243151 observed at 25kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab243151 Anti-BAK1 antibody [7D10] was shown to react with BAK1 in HeLa cells in Western blot. Loss of signal was observed when BAK1 knockout sample was used. Wild-type and BAK1 knockout samples were subjected to SDS-PAGE.

Lanes 3-4 : ab243151 Anti-BAK1 antibody [7D10] was shown to react with BAK1 in HAP1 cells in Western blot. Loss of signal was observed when BAK1 knockout sample was used. Wild-type and BAK1 knockout samples were subjected to SDS-PAGE.

ab243151 and Anti-GAPDH antibody [6C5] (ab8245) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat Anti-Rat IgG H&L (IRDye^® 800CW) (ab253031) and Goat Anti-Mouse IgG H&L (IRDye^® 680RD) (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bak antibody [7D10] (<a href='/en-us/products/primary-antibodies/bak-antibody-7d10-ab243151'>ab243151</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

BAK1 knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate at 20 µg

Lane 4:

BAK1 knockout HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rat IgG H&L (IRDye® 800CW) (ab253031) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false