PD-1/CD279 Monoclonal antibody, PBS Only

PD-1/CD279 Monoclonal Antibody for WB, IF, FC, IHC, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, rat, mouse

Applications

WB, IF, FC, IHC, ELISA

Conjugate

Unconjugated

CloneNo.

4H4D1

Cat no : 66220-1-PBS

Synonyms

CD279, hPD 1, hPD l, PD1, PD-1, PD-1/CD279, PDCD1, programmed cell death 1, Protein PD 1, SLEB2

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Validation Data Gallery

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  WB analysis using 66220-1-Ig (same clone as 66220-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Lamin B1 Monoclonal antibody (HRP-66095) as loading control. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  WB analysis using 66220-1-Ig (same clone as 66220-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Lamin B1 Monoclonal antibody (HRP-66095) as loading control. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  WB analysis of mouse thymus using 66220-1-Ig (same clone as 66220-1-PBS)

  mouse thymus tissue was subjected to SDS PAGE followed by western blot with 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  WB analysis of rat spleen using 66220-1-Ig (same clone as 66220-1-PBS)

  rat spleen tissue were subjected to SDS PAGE followed by western blot with 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  WB analysis of human lymph node using 66220-1-Ig (same clone as 66220-1-PBS)

  human lymph node tissue were subjected to SDS PAGE followed by western blot with 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:4000 (under 4x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

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  IHC staining of human lymphoma using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human angioimmunoblastic T-cell lymphoma tissue slide using 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:200 (under 10x lens). . Heat mediated antigen retrieved with Tris-EDTA buffer, pH9.0. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human lymphoma using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human angioimmunoblastic T-cell lymphoma tissue slide using 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:200 (under 40x lens). . Heat mediated antigen retrieved with Tris-EDTA buffer, pH9.0. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieved with Tris-EDTA buffer, pH9.0. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieved with Tris-EDTA buffer, pH9.0. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:2000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66220-1-Ig (PD-1/CD279 antibody) at dilution of 1:2000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using PD-1/CD279 antibody (66220-1-Ig, Clone: 4H4D1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using PD-1/CD279 mouse mAb (66220-1-Ig) at dilution of 1:50 and CD20 rabbit pAb (24828-1-AP) at dilution of 1:50, further stained with Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L) for 66220-1-Ig, and Alexa Fluor 594-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) for 24828-1-AP. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human lymphoma using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded human angioimmunoblastic T-cell lymphoma tissue using PD-1/CD279 antibody (66220-1-Ig, Clone: 4H4D1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human lymphoma using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded human angioimmunoblastic T-cell lymphoma tissue using PD-1/CD279 antibody (66220-1-Ig, Clone: 4H4D1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human tonsillitis using 66220-1-Ig (same clone as 66220-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using PD-1/CD279 mouse mAb (66220-1-Ig) at dilution of 1:50 and CD20 rabbit pAb (24828-1-AP) at dilution of 1:50, further stained with Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L) for 66220-1-Ig, and Alexa Fluor 594-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) for 24828-1-AP. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

• 

  FC experiment of MOLT-4 using 66220-1-Ig (same clone as 66220-1-PBS)

  1X10^6 unstimulated (dashed line) or PMA and ionomycin treated (red) MOLT-4 cells were surface stained with 0.2 ug Anti-Human PD-1/CD279 (66220-1-Ig, Clone: 4H4D1) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000, or 0.2 ug isotype control antibody (blue, solid line). Cells were not fixed. This data was developed using the same antibody clone with 66220-1-PBS in a different storage buffer formulation.

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Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

66220-1-PBS targets PD-1/CD279 in WB, IF, FC, IHC, ELISA applications and shows reactivity with human, rat, mouse samples.

• Tested Reactivity: human, rat, mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: PD-1/CD279 fusion protein Ag12470
• Full Name: programmed cell death 1
• Calculated Molecular Weight: 288 aa, 32 kDa
• Observed Molecular Weight: 32 kDa, 47-55 kDa
• GenBank Accession Number: BC074740
• Gene Symbol: PDCD1
• Gene ID (NCBI): 5133
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS only
• Storage Conditions: Store at -20°C. Stable for one year after shipment.

Background Information

Programmed cell death 1 (PD-1, also known as CD279) is an immunoinhibitory receptor that belongs to the CD28/CTLA-4 subfamily of the Ig superfamily. It is a 288 amino acid (aa) type I transmembrane protein composed of one Ig superfamily domain, a stalk, a transmembrane domain, and an intracellular domain containing an immunoreceptor tyrosine-based inhibitory motif (ITIM) as well as an immunoreceptor tyrosine-based switch motif (ITSM) (PMID: 18173375). PD-1 is expressed during thymic development and is induced in a variety of hematopoietic cells in the periphery by antigen receptor signaling and cytokines (PMID: 20636820). Engagement of PD-1 by its ligands PD-L1 or PD-L2 transduces a signal that inhibits T-cell proliferation, cytokine production, and cytolytic function (PMID: 19426218). It is critical for the regulation of T cell function during immunity and tolerance. Blockade of PD-1 can overcome immune resistance and also has been shown to have antitumor activity (PMID: 22658127; 23169436). The calculated molecular weight of PD-1 is 32 kDa. It has been reported that PD-1 is heavily glycosylated and migrates with an apparent molecular mass of 47-55 kDa on SDS-PAGE (PMID: 8671665; 17640856; 17003438).