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How To Choose An Anti-PD-1 Antibody For Your Research

PD-1 Introduction

Programmed death-1 (PD-1) is an inhibitory receptor expressed on T-cells that promotes apoptosis of inflammatory T-cells and inhibits apoptosis of anti-inflammatory regulatory T-cells, thereby promoting self-tolerance and preventing autoimmune diseases. To evade immune surveillance, tumor cells often exploit this system by over-expressing PD-L1, the ligand which binds to and activates PD-1. Anti-PD-1 monoclonal antibodies are used to block the binding of PD-L1 to PD-1 which allows the immune system to find and kill tumor cells.

Anti-PD-1 Antibody Reactivity

The first step in deciding which anti-PD-1 antibody fits your research needs is to identify which species reactivity is needed for your experimental model. Bio X Cell offers anti-mouse and anti-human PD-1 antibodies. Which to choose is based on your experimental model. If you are using mice expressing mouse PD-1 you want to choose an anti-mouse PD-1 antibody. If you are using human cells in culture or humanized mice expressing human PD-1 then you want to choose an anti-human PD-1 antibody.

Anti-mouse PD-1 Antibody Clones

Bio X Cell offers three distinct anti-mouse PD-1 antibody clones: RMP1-14, 29F.1A12, and J43. All three antibodies function through the same mechanism ā€“ they bind to PD-1 and sterically block the binding of PD-1 to PD-1 ligands, thereby blocking PD-1 signaling (as shown in the image above). These antibodies do not function through depleting PD-1 expressing cells.

All three antibodies are ideally suited for in vivo blocking of PD-1 signaling in mouse models and have extensive multi-year publication records backing this application. The differences between these antibodies lie in their PD-1 blocking capacities, their listed reported (published) applications, their host and isotype, and their origins.

With respect to their PD-1 blocking capacities, a recent publication titled ā€œA Comparison of Murine PD-1 and PD-L1 Monoclonal Antibodiesā€ compares the RMP1-14 and 29F.1A12 clones. The authors found that the 29F.1A12 clone has approximately 100-fold higher PD-1 blocking affinity than the RMP1-14 clone. The authors further conclude that 29F.1A12 most closely models human PD-1 therapeutic antibodies, which have a high affinity and blocking ability. Further details on this study can be found here:

 

Comparing PD-1 and PD-L1 Mouse Monoclonal Antibodies

Among these anti-PD-1 antibodies, the RMP1-14 clone has the most extensive publication record for in vivo blocking of PD-1 signaling. Importantly, however, the reported applications for the RMP1-14 antibody are limited to in vivo blocking of PD-1/PD-L signaling only.

Alternatively, the 29F.1A12 clone also has an extensive publication record for in vivo PD-1 blocking but can also be used for in vitro PD-1 neutralization, western blotting, immunohistochemistry, immunofluorescence, and flow cytometry. The J43 clone fits a similar description. This can be advantageous for researchers who wish to run in vitro or diagnostic experiments in parallel with in vivo experiments.

The next step in making the antibody selection that is best for your research is to search the publication record for each clone and find an example of published data that uses a similar experimental approach to your own. There may be one clone that is more often used for your specific mouse model and experimental set-up.

Bio X Cell provides a list of selected references for all our antibodies which can be found on each product page. Additional references can be found using search engines such as Google Scholar and PubMed. Searching for the clone name of the antibody in combination with experiment-specific search terms, for example, ā€œRMP1-14 MC38 BALB/cā€, is the best approach to finding applicable references. These references will help you select the antibody that best fits your research.

Detailed Comparison of anti-mouse PD-1 Antibody Clones

Clone RMP1-14 29F.1A12 J43
InVivoPlusā„¢ Catalog no. BP0146 BP0273 BP0033-2
Isotype Rat IgG2a, Īŗ Rat IgG2a Armenian Hamster IgG
First Citation In 2003 by Takanori Kanai et al In 2003 by Spencer C. Liang et al In 1996 by Yasutoshi Agata et al
Reported Applications in vivo blocking of PD-1/PD-L signaling
  • in vivo blocking of PD-1/PD-L signaling
  • in vitro PD-1 neutralization
  • Immunohistochemistry (frozen)
  • Immunofluorescence
  • Western blot
  • Flow cytometry
  • in vivo blocking of PD-1/PD-L signaling
  • in vitro PD-1 neutralization
  • Western blot
Immunogen Syrian Hamster BKH cells transfected with mouse PD-1 cDNA Recombinant PD-1-Ig fusion protein Syrian Hamster BKH cells transfected with mouse PD-1 cDNA
Purity >95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE
Stabilizers or Preservatives Free Free Free
Size 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more
Availability In stock In stock In stock
Recommended Isotype Control InVivoPlusā„¢ rat IgG2a isotype control, anti-trinitrophenol InVivoPlusā„¢ rat IgG2a isotype control, anti-trinitrophenol InVivoPlusā„¢ polyclonal Armenian hamster IgG
Recommended Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 6.5 Dilution Buffer

All of Bio X Cellā€™s InVivoPlusā„¢ antibodies, including these anti-PD-1 clones, are rigorously tested to ensure that they are murine pathogen free, have an endotoxin level of <1EU/mg, and a protein aggregation level of <5%. These clones are also available in our cost effective InVivoMAbā„¢ format. These alternatives are: RMP1-14 (BE0146), 29F.1A12 (BE0273), and J43 (BE0033-2). A detailed explanation of the differences between our InVivoPlusā„¢ and InVivoMAbā„¢ formats can be viewed at the link below.

InVivoPlusā„¢ vs. InVivoMAbā„¢

Recombinant anti-mouse PD-1 Antibody Clones

Bio X Cell also offers recombinant chimeric versions of our three anti-mouse PD-1 antibody clones. RecombiMAbā„¢ antibodies are provided as chimeric antibodies with mouse constant regions instead of the typical rat or hamster constant regions. This ensures reduced immunogenicity in sensitive mouse models. Select RecombiMAbā„¢ antibodies are also available in multiple Fc silenced versions. These Fc silenced versions have mutations in the Fc fragment of the antibody rendering them unable to bind to endogenous FcĪ³ receptors.

Learn More About RecombiMAbā„¢ Antibodies

Detailed Comparison of Recombinant anti-mouse PD-1 Antibody Clones

Clone RMP1-14-CP153 RMP1-14-CP151 RMP1-14-CP162 29F.1A12ā„¢-CP159 RMP1-14-CP157 29F.1A12ā„¢-CP155
InVivoPlusā„¢ Catalog no. CP153 CP151 CP162 CP159 CP157 CP155
Isotype Mouse IgG2a, Īŗ Mouse IgG2a, Īŗ Mouse IgG1, Īŗ Mouse IgG2a, Īŗ Mouse IgG2a, Īŗ Mouse IgG2a, Īŗ
Mutations LALA-PG D265A None None None LALA-PG
Reported Applications in vivo blocking of PD-1/PD-L signaling* *Reported for the original rat IgG2a RMP1-14 antibody
  • in vivo blocking of PD-1/PD-L signaling* *Reported for the original rat IgG2a RMP1-14 antibody
  • in vivo blocking of PD-1/PD-L signaling* *Reported for the original rat IgG2a RMP1-14 antibody
  • in vivo blocking of PD-1/PD-L signaling*
  • in vitro PD-1 neutralization*
  • Immunohistochemistry (frozen)*
  • Immunofluorescence*
  • Western blot*
  • Flow cytometry*
  • *Reported for the original rat IgG2a 29F.1A12 antibody
  • in vivo blocking of PD-1/PD-L signaling*
  • *Reported for the original rat IgG2a RMP1-14 antibody
  • in vivo blocking of PD-1/PD-L signaling*
  • in vitro PD-1 neutralization*
  • Immunohistochemistry (frozen)*
  • Immunofluorescence*
  • Western blot*
  • Flow cytometry*
  • *Reported for the original rat IgG2a 29F.1A12 antibody
Immunogen Syrian Hamster BKH cells transfected with mouse PD-1 cDNA Syrian Hamster BKH cells transfected with mouse PD-1 cDNA Syrian Hamster BKH cells transfected with mouse PD-1 cDNA Recombinant PD-1-Ig fusion protein Syrian Hamster BKH cells transfected with mouse PD-1 cDNA Recombinant PD-1-Ig fusion protein
Purity >95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE
Stabilizers or Preservatives Free Free Free Free Free Free
Size 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more
Availability In stock In stock In stock In stock In stock In stock
Recommended Isotype Control InVivoRecombiMAb mouse IgG2a (D265A) isotype control, anti-hen egg lysozyme InVivoRecombiMAb mouse IgG2a (D265A) isotype control, anti-hen egg lysozyme InVivoInVivoPlus mouse IgG1 isotype control, unknown specificity InVivoInVivoPlus mouse IgG2a isotype control, unknown specificity InVivoInVivoPlus mouse IgG2a isotype control, unknown specificity InVivoRecombiMAb mouse IgG2a (LALA-PG) isotype control, unknown specificity
Recommended Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer

Anti-human PD-1 Antibody Clones

Bio X Cell also offers multiple anti-human PD-1 antibodies for use in humanized mouse models or immunoassays. These include clones J116 and J110 and Pembrolizumab and Nivolumab biosimilar antibodies.

Detailed Comparison of anti-human PD-1 Antibody Clones

Clone Nivolumab Pembrolizumab J116 407.6G12 J110
InVivoPlusā„¢ Catalog no. SIM0003 SIM0010 BE0188 BE0387 BE0193
Isotype Human IgG4 Human IgG4 Mouse IgG1, Īŗ Mouse IgG2a, Īŗ Mouse IgG1, Īŗ
Reported Applications
  • Blocking of PD-1/PD-L signaling
  • Western Blot
  • Immunohistochemistry
  • Flow cytometry
  • Blocking of PD-1/PD-L signaling
  • Functional assays
  • in vitro PD-1 neutralization
  • in vivo PD-1 blockade in humanized mice
  • Western Blot
  • Flow Cytometry
  • in vivo PD-1 blockade in humanized mice
  • Flow Cytometry
Immunogen Human PD-1 Human PD-1 Not available or unknown Human PD-1 p248Tyr peptide conjugated to KLH Not available or unknown
Purity >95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE 		>95%
Determined by SDS-PAGE
Stabilizers or Preservatives Free Free Free Free Free
Size 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more 5, 25, 50, 100 mg and more
Availability In stock In stock In stock In stock In stock
Recommended Isotype Control InVivoRecombiMAb human IgG4 (S228P) isotype control, anti-hen egg lysozyme InVivoRecombiMAb human IgG4 (S228P) isotype control, anti-hen egg lysozyme InVivoInVivoMAb mouse IgG1 isotype control, unknown specificity InVivoInVivoMAb mouse IgG2a isotype control, unknown specificity InVivoInVivoMAb mouse IgG1 isotype control, unknown specificity
Recommended Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer InVivoPureā„¢ pH 7.0 Dilution Buffer