Anti-PD1 antibody [NAT105]

5 product images

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] (ab234444)

  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.

• 

  Flow Cytometry (Intracellular) - Anti-PD1 antibody [NAT105] (ab234444)

  Intracellular 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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• 

  Western blot - Anti-PD1 antibody [NAT105] (ab234444)

  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.

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

  Lane 1: Untreated MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate at 10 µg

  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 at 10 µg

  Secondary

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

  Predicted band size: 32 kDa

  Exposure time: 10s

• 

  Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [NAT105] (ab234444)

  ab234444 staining PD1 in MOLT4 treated with Ionomycin (500 ng/ml, 24 h) and PMA (10 ng/ml, 24 h) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab234444 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• 

  Flow Cytometry - Anti-PD1 antibody [NAT105] (ab234444)

  Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab234444 (right) or Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab234444 or Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (1x 106 in 100 µl at 5.0 μg/ml (1/516)) for 30min on ice. The cells were simultaneously stained with CD3.
  
  The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
  
  Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.