Anti-RPA32/RPA2 antibody [EPR2877Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1 : 50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 μg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/50 dilution (2.0 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). 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76420 (unpurified) at 1/250 dilution.

• IP

Unknown

Immunoprecipitation - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

ab76420 (purified) at 1/20 dilution (0.5ug) immunoprecipitating RPA32/RPA2 in HeLa whole cell lysate. HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab76420 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76420 in HeLa whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420)

Predicted band size: 29 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 μg/ml). Heat mediated antigen retrieval was performed ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Intracellular Flow Cytometry analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 μg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Intracellular Flow Cytometry analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 μg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1 : 50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1 : 50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/1000 dilution

Lane 1:

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

Lane 2:

C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 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: 29 kDa

Observed band size: 29 kDa

false

• WB

Unknown

Western blot - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HUVEC lysate at 10 µg

Lane 3:

NIH 3T3 cell lysate at 10 µg

Lane 4:

C6 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/1000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

false

• WB

Lab

Western blot - Anti-RPA32/RPA2 antibody [EPR2877Y] (AB76420)

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

The molecular weight of RPA32/RPA2 is upshifted in response to camptothecin treatment (PMID : 17928296).

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.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-phospho-s33-antibody-epr29612-551-ab325750'>ab325750</a>) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) fresh whole cell lysate at 50 µg

Lane 2:

NIH/3T3 treated with 1 μM camptothecin for 3 h fresh 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: 32 kDa,36 kDa,30 kDa

true

Exposure time: 70s