Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PD1 with ab234444 at 1/50 dilution, followed by Rabbit Anti-Mouse IgG + Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on T cells of human tonsil germinal center is observed. Performed on a Leica Biosystems BOND instrument. Counter stained with hematoxylin.

Ab234444 staining PD1 in the MOLT-4 (human lymphoblastic leukemia T lymphoblast) treated with Ionomycin (500 ng/ml, 24 h) and PMA (10 ng/ml, 24 h) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/50). ab150117 anti-mouse IgG H&L (Alexa Fluor® 488) preadsorbed (1/100) was used as the secondary antibody. 

ab179504 Anti-beta IV Tubulin (1/200) was used as a counter stain, detected by ab150080 AlexaFluor®594 Goat anti- Rabbit secondary (1/500)

DAPI was used as a Nuclear counter stain.

Flow cytometric analysis of MOLT-4 (human lymphoblastic leukemia cell line) cell line treated with ionomycin (500 ng/ml, 24h) and PMA (10 ng/ml, 24h) labeling PD1 with ab234444 at 1/1000 dilution (red) and an untreated control (green) compared with a Mouse monoclonal IgG1 (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

All lanes : Anti-PD1 antibody [NAT105] (ab234444) at 1/1000 dilution

Lane 1 : Untreated MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate
Lane 2 : MOLT-4 (human lymphoblastic leukemia cell line) treated with 500 ng/ml ionomycin and 10 ng/ml phorbol-12-myristate-13-acetate (PMA) for 24 hours, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-Mouse IgG (H+L) Peroxidase Peroxidase-Conjugated at 1/5000 dilution

Predicted band size: 32 kDa


Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 26318293)

We recommend that the customer optimizes the blotting conditions for themselves.