Anti-p53R2 antibody [EPR8816] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (AB249090)

This data was developed using ab154194, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Wild-type HeLa (Human cervical adenocarcinoma epithelial cell) cell pellet (A) and RRM2B KO HeLa cell pellet (B) labelling p53R2 with ab154194 at 1 : 2500 (0.044 μg/ml). Positive staining on (A) wild-type HeLa cell pellet, no staining on (B) RRM2B KO HeLa A431 cell pellet. The primary antibody was incubated for 30 mins at room temperature. Secondary antibody was a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (AB249090)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling p53R2 with purified ab154194 at 1/500 dilution (0.22 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

• IP

Unknown

Immunoprecipitation - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (AB249090)

ab154194 (purified ) at 1/20 dilution (0.5ug) immunoprecipitating p53R2 in MCF7 whole cell lysate.

Lane 1 (input) : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab154194 & MCF7 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab154194 in MCF7 whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

All lanes:

Immunoprecipitation - Anti-p53R2 antibody [EPR8816] (<a href='/en-us/products/primary-antibodies/p53r2-antibody-epr8816-ab154194'>ab154194</a>)

Predicted band size: 40 kDa

false

• WB

Lab

Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (AB249090)

This data was developed using the same antibody clone in a different buffer formulation (ab154194).

Lanes 1- 2 : Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab154194 was shown to react with p53R2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261769 (knockout cell lysate ab257215) was used. Wild-type HeLa and RRM2B knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-p53R2 antibody [EPR8816] (<a href='/en-us/products/primary-antibodies/p53r2-antibody-epr8816-ab154194'>ab154194</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

RRM2B knockout HeLa cell lysate at 40 µg

Lane 2:

Western blot - Human RRM2B (p53R2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rrm2b-p53r2-knockout-hela-cell-line-ab261769'>ab261769</a>)

Predicted band size: 40 kDa

Observed band size: 40 kDa

false

• WB

Lab

Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (AB249090)

This data was developed using the same antibody clone in a different buffer formulation (ab154194).

Lanes 1- 2 : Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab154194 was shown to react with p53R2 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266897 (knockout cell lysate ab257216) was used. Wild-type HCT116 and RRM2B knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-p53R2 antibody [EPR8816] (<a href='/en-us/products/primary-antibodies/p53r2-antibody-epr8816-ab154194'>ab154194</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

RRM2B knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human RRM2B (p53R2) knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-rrm2b-p53r2-knockout-hct116-cell-line-ab266897'>ab266897</a>)

Predicted band size: 40 kDa

Observed band size: 40 kDa

false