Anti-XPD antibody [4G2-2A6]

• Flow Cyt

Unknown

Flow Cytometry - Anti-XPD antibody [4G2-2A6] (AB54676)

Overlay histogram showing HeLa cells stained with ab54676 (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 (ab54676, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

• IP

Unknown

Immunoprecipitation - Anti-XPD antibody [4G2-2A6] (AB54676)

XPD was immunoprecipitated using 0.5mg Hela whole cell extract, 10μg of Mouse monoclonal to XPD and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab54676.
Secondary : Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band : 150kDa : SMC1; Non specific - 41 and 42kDa : We are unsure as to the identity of this extra band.

This image was generated using the ascites version of the product.

All lanes:

Immunoprecipitation - Anti-XPD antibody [4G2-2A6] (ab54676)

Predicted band size: 87 kDa

false

• WB

Unknown

Western blot - Anti-XPD antibody [4G2-2A6] (AB54676)

XPD antibody (ab54676) at 1ug/lane + HeLa cell lysate at 25ug/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-XPD antibody [4G2-2A6] (ab54676)

Predicted band size: 87 kDa

false