Recombinant Anti-CCR7 antibody [Y59] - Low endotoxin, Azide free (ab221209)

ab32527 (purified) at 1/20 immunoprecipitating CCR7 in MCF7 (human breast adenocarcinoma cell line) whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) was used for detection at 1/10,000 dilution. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32527).

Immunocytochemistry/Immunofluorescence analysis of Jurkat (human  T cell leukemia cell line from peripheral blood) cells labelling CCR7 with purified ab32527 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32527).

Immunocytochemistry/Immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling CCR7 with unpurified ab32527 at 1/1000. Cells were fixed with 4% PFA fixed (10 min) and permeabilized and blocked with 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with the antibody (ab32527, 1µg/ml) overnight at +4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (shown in green). Alexa Fluor^® 594 WGA was used to label plasma membranes (shown in red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32527).