Anti-RPA70 antibody [EPR3472] - BSA and Azide free

• ICC/IF

AbReview46343****

Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Unpurified ab79398 staining RPA70 in U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 in PBS and blocked with 2% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/500 in PBS + 0.5% Tween-20) for 2 hours at 25°C. A Cy3^®-conjugated goat anti-rabbit IgG monoclonal (1/250) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

This image is courtesy of an Abreview submitted by Remi Buisson.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical squamous cell carcinoma labelling RPA70 with unpurified ab79398 at a dilution of 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Overlay histogram showing HeLa cells stained with unpurified ab79398 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab79398, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling RPA70 with purified ab79398 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Immunocytochemistry/Immunofluorescence analysis of A549 cells labelling RPA70 with purified ab79398 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.

Control 1 : primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

Intracellular Flow Cytometry analysis of HeLa cells labelling RPA70 with purified ab79398 at 1/80 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

• IP

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Immunoprecipitation - Anti-RPA70 antibody [EPR3472] - BSA and Azide free (AB239890)

ab79398 (purified) at 1/20 immunoprecipitating RPA70 in HeLa whole cell lysate.

Lane 1 (input) : HeLa whole cell lysate (10µg)

Lane 2 (+) : ab79398 + HeLa whole cell lysate (10µg).

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab79398 in HeLa whole cell lysate.

For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

All lanes:

Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (<a href='/en-us/products/primary-antibodies/rpa70-antibody-epr3472-ab79398'>ab79398</a>)

Predicted band size: 68 kDa

Observed band size: 70 kDa

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