Anti-XRCC1 antibody (ab47920)

Western blot - XRCC1 antibody (ab47920)

All lanes : Anti-XRCC1 antibody (ab47920) at 1/250 dilution

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 70 kDa
Observed band size : 80 kDa (why is the actual band size different from the predicted?)

The band seen at 80 kDa is consistent with the banding pattern observed for other commercially available antibodies to XRCC1.

Immunocytochemistry/ Immunofluorescence - XRCC1 antibody (ab47920)

ICC/IF image of ab47920 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in PBS-Tween20 0.1% (20 min) and incubated with the antibody (ab47920, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). Antibodies were diluted in 1% BSA, 1% normal goat serum in PBS-Tween.

Immunocytochemistry/ Immunofluorescence - XRCC1 antibody (ab47920)

ab47920 (1/200) staining XRCC1 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - XRCC1 antibody (ab47920)

ab47920 staining XRCC1 in human tissue sections by immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in citrate buffer pH6.0 and blocking for 15 minutes at 20°C (5 minutes for peroxidase block and 10 minutes for protein block). The primary antibody was diluted 1/50 and incubated with sample for 45 minutes at 20°C. A HRP-conjugated goat polyclonal to rabbit IgG was used undiluted as secondary. 

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.