Anti-XPF antibody [EPR25350-148] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-XPF antibody [EPR25350-148] - BSA and Azide free (AB290740)

This data was developed using ab290725, the same antibody clone in a different buffer formulation.

XPF was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug with ab290725 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290725 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug

Lane 2 : ab290725 IP in 293T whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab290725 in 293T whole cell lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-XPF antibody [EPR25350-148] (<a href='/en-us/products/primary-antibodies/xpf-antibody-epr25350-148-ab290725'>ab290725</a>)

Predicted band size: 104 kDa

false

• WB

Lab

Western blot - Anti-XPF antibody [EPR25350-148] - BSA and Azide free (AB290740)

This data was developed using ab290725, the same antibody clone in a different buffer formulation.

Blocking / Diluent buffer and concentration : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1-3 : Merged signal (red and green). Green - ab290725 observed at 104 kDa. Red - loading control ab8245 observed at 36 kDa.

ab290725 Anti-ERCC4 antibody [EPR25350-148] was shown to specifically react with ERCC4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line was used. Wild-type and ERCC4 knockout samples were subjected to SDS-PAGE. ab290725 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-XPF antibody [EPR25350-148] (<a href='/en-us/products/primary-antibodies/xpf-antibody-epr25350-148-ab290725'>ab290725</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate. at 20 µg

Lane 2:

ERCC4 knockout HeLa whole cell lysate. at 20 µg

Lane 3:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 104 kDa

Observed band size: 104 kDa

false

• WB

Lab

Western blot - Anti-XPF antibody [EPR25350-148] - BSA and Azide free (AB290740)

This data was developed using ab290725, the same antibody clone in a different buffer formulation.

Blocking / Dilution buffer and concentration : 5% NFDM/TBST

Lysates should be made freshly and used in WB immediately to minimize protein degradation.

All lanes:

Western blot - Anti-XPF antibody [EPR25350-148] (<a href='/en-us/products/primary-antibodies/xpf-antibody-epr25350-148-ab290725'>ab290725</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate. at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate. at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) 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 1/20000 dilution

Predicted band size: 104 kDa

Observed band size: 104 kDa

false

Exposure time: 15s