Recombinant Anti-Midkine antibody [EP1143Y] - BSA and Azide free (ab236781)

ab52637 (purified) at 1:20 dilution (2ug) immunoprecipitating Midkine in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate.

Lane 1 (input): SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate 10ug

Lane 2 (+): ab52637 & SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52637 in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:5000 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 (ab52637).

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Midkine with purified ab52637 at 1:100 dilution (10.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52637).

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Midkine with purified ab52637 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52637). 

Human liver carcinoma stained with unpurified ab52637 at 1/100 - 1/250 dilution

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52637).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

HeLa cells stained with unpurified ab52637 at 1/250 - 1/500 dilution

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52637).