Antibodies, Proteins, Kits and Reagents for Life Science

341 740 ₸ Product size

100 µl 341 740 ₸ 10 µl 120 614 ₸

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Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [NAT105] to PD1 - Chimeric
Suitable for: WB, IHC-P, Flow Cyt, IP
Reacts with: Human

Product name

Anti-PD1 antibody [NAT105] - Chimeric
See all PD1 primary antibodies
Description

Rabbit monoclonal [NAT105] to PD1 - Chimeric
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Tissue, cells or virus corresponding to Human PD1. Specifically, human YT cells (NK-like leukemia cell line) that express PD1.
Database link: Q15116
Positive control
- WB: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours, whole cell lysate; Human tonsil lysate. IHC-P: Human tonsil and bladder carcinoma tissues. Flow Cyt: MOLT-4 cells treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours. IP: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.
General notes

This rabbit antibody has been engineered from the mouse monoclonal parent antibody Anti-PD1 antibody [NAT105] (ab52587). By necessity, some mouse sequence is retained as part of the variable domain.

We recommend using an Fc-directed, preadsorbed secondary antibody if multiplexing with mouse monoclonals. For example, ab150097 (AF488), ab150100 (AF594), ab150099 (AF647) or ab98467 (HRP).

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

NAT105
Isotype

IgG
Research areas
- Cancer
- Cell Death
- Apoptosis
- Other

Flow Cytometry - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

Flow cytometric analysis of MOLT-4 (human lymphoblastic leukemia cell line) cell line treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours (Red) or Untreated control (Green) labeling PD1 with ab216352 at 1/600 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PD1 with ab216352 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on T cells of human tonsil germinal center (PMID: 20087161; PMID: 17640856) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.

Perform antigen retrieval using Universal HIER antigen retrieval reagents (10X) (ab208572).
Immunoprecipitation - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

PD1 was immunoprecipitated from 0.35 mg of 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 with ab216352 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216352 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution

Lane 1: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate 10 µg (Input).

Lane 2: ab216352 IP in MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216352 in MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.

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

Exposure time: 40 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

Immunohistochemical analysis of paraffin-embedded human bladder carcinoma tissue labeling PD1 with ab216352 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on lymphocytes in human bladder carcinoma (PMID: 20087161, PMID: 17640856) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Perform antigen retrieval using Universal HIER antigen retrieval reagents (10X) (ab208572).
Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

Anti-PD1 antibody [NAT105] - Chimeric (ab216352) at 1/1000 dilution + Human tonsil lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 32 kDa
Observed band size: 50-55 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).
Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

All lanes : Anti-PD1 antibody [NAT105] - Chimeric (ab216352) at 1/1000 dilution

Lane 1 : Untreated MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate
Lane 2 : MOLT-4 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-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 32 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?

Exposure time: 81 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).
Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

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