Recombinant Anti-ADRM1/ARM-1 antibody [EPR11449(B)] (ab157185)

Immunocytochemistry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling ADRM1/ARM-1 with Purified ab157185 at 1:1000 dilution (0.6 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A]+H21:L21 - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric cancer tissue sections labeling ADRM1/ARM-1 with Purified ab157185 at 1:9000 (0.07 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling ADRM1/ARM-1 with Purified ab157185 at 1:9000 (0.07 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling ADRM1/ARM-1 with Purified ab157185 at 1:60 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Lanes 1-4: Merged signal (red and green). Green - ab157185 observed at 42 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab157185 Anti-ADRM1/ARM-1 antibody [EPR11449(B)] was shown to specifically react with ADRM1/ARM-1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266765 (knockout cell lysate ab257816) was used. Wild-type and ADRM1/ARM-1 knockout samples were subjected to SDS-PAGE. ab157185 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 20000 dilution for 1 hour at room temperature before imaging.

ADRM1/ARM-1 was immunoprecipitated from 0.35 mg Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 µg with 157185 at 1/100 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 µg

Lane 2: ab157185 IP in Raji whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab157185 in Raji whole cell lysate

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