Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling BRCA2 with ab322104 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab322104 anti-BRCA2 antibody [EPR26146-81] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human invasive lobular breast carcinoma labelling BRCA2 with ab322104 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab322104 anti-BRCA2 antibody [EPR26146-81] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human ductal breast carcinoma labelling BRCA2 with ab322104 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab322104 anti-BRCA2 antibody [EPR26146-81] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling BRCA2 with ab322104 at 1/500 (1.006 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Low expression tissue : almost no staining on human lung (PMID : 9133444). The section was incubated with ab322104 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 LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling BRCA2 with ab322104 at 1/500 (1.006 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab322104 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 LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling BRCA2 with ab322104 at 1/500 (1.006 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human brest carcinoma (PMID : 15933754). The section was incubated with ab322104 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 LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

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

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW band at approximately 130 kDa is unknown.

The band beneath the target band (200 kDa) is expected to be degradation product.

In Western blot Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-BRCA2 antibody [EPR26146-81] (<a href='/en-us/products/primary-antibodies/brca2-antibody-epr26146-81-ab322104'>ab322104</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

Hela transfected with siRNA specifically targeting BRCA2 whole cell lysate at 50 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

Observed band size: 200 kDa,384 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-BRCA2 antibody [EPR26146-81] - BSA and Azide free (AB324804)

This data was developed using ab322104, the same antibody clone in a different buffer formulation.

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

Capan-1 cells have one copy of BRCA2 gene missing and a mutation in the remaining copy resulting in a truncation of the protein (180kDa).

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The band beneath the target band (250 kDa) is likely to be degradation product.

The identity of the lower MW band at approximately 130 kDa is unknown.

In Western blot Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-BRCA2 antibody [EPR26146-81] (<a href='/en-us/products/primary-antibodies/brca2-antibody-epr26146-81-ab322104'>ab322104</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 100 µg

Lane 2:

Capan-1 (human pancreatic adenocarcinoma epithelial cell) whole cell lysate at 100 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

Observed band size: 180 kDa,250 kDa,384 kDa,124 kDa

false

Exposure time: 180s