Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling XRCC4 with ab317697 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human tonsil.
The section was incubated with ab317697 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-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human endometrioid carcinoma tissue labeling XRCC4 with ab317697 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human endometrioid carcinoma.
The section was incubated with ab317697 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-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling XRCC4 with ab317697 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human breast.
The section was incubated with ab317697 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

• IP

Supplier Data

Immunoprecipitation - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

XRCC4 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab317697 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317697 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab317697 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317697 in HeLa whole cell lysate

All lanes:

Immunoprecipitation - Anti-XRCC4 antibody [EPR28958-63] (<a href='/en-us/products/primary-antibodies/xrcc4-antibody-epr28958-63-ab317697'>ab317697</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 50s

• WB

Supplier Data

Western blot - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-XRCC4 antibody [EPR28958-63] (<a href='/en-us/products/primary-antibodies/xrcc4-antibody-epr28958-63-ab317697'>ab317697</a>) at 1/1000 dilution

All lanes:

Human tonsil tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 35 kDa,55 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-XRCC4 antibody [EPR28958-63] (<a href='/en-us/products/primary-antibodies/xrcc4-antibody-epr28958-63-ab317697'>ab317697</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti XRCC4 whole cell lysate at 20 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 1/100000 dilution

Observed band size: 35 kDa,55 kDa,124 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

The expression of caspase cleaved XRCC4 is upregulated upon induction of apoptosis (PMID : 33725486).

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

All lanes:

Western blot - Anti-XRCC4 antibody [EPR28958-63] (<a href='/en-us/products/primary-antibodies/xrcc4-antibody-epr28958-63-ab317697'>ab317697</a>) at 1/1000 dilution

Lane 1:

Untreated heLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 2uM staurosporine for 3h whole cell lysate at 20 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 1/100000 dilution

Observed band size: 35 kDa,55 kDa,124 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-XRCC4 antibody [EPR28958-63] - BSA and Azide free (AB317698)

This data was developed using ab317697, the same antibody clone in a different buffer formulation.

In Lane 1-Lane 4, lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-XRCC4 antibody [EPR28958-63] (<a href='/en-us/products/primary-antibodies/xrcc4-antibody-epr28958-63-ab317697'>ab317697</a>) at 1/1000 dilution

Lanes 1 and 6:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 5:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 7:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 55 kDa

false

Exposure time: 15s