Recombinant Anti-PD1 antibody [EPR23119-111] (ab243644)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23119-111] to PD1
- Suitable for: mIHC, IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PD1 antibody [EPR23119-111]
See all PD1 primary antibodies -
Description
Rabbit monoclonal [EPR23119-111] to PD1 -
Host species
Rabbit -
Tested applications
Suitable for: mIHC, IHC-P, WBmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Untreated MOLT-4, MOLT-4 treated with 500ng/ml Ionomycin and 10ng/ml Phorbol-12-myristate-13-acetate and Human tonsil, Human tonsil treated with PNGase F lysates. IHC-P: Human tonsil and Human Hodgkin's lymphoma, Human endometrial carcinoma tissues. mIHC: Human tonsil and Human spleen tissue
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23119-111 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab243644 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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mIHC |
1/500.
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IHC-P |
1/500.
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WB |
1/1000. Predicted molecular weight: 32 kDa.
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Notes |
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mIHC
1/500. |
IHC-P
1/500. |
WB
1/1000. Predicted molecular weight: 32 kDa. |
Target
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Function
Possible cell death inducer, in association with other factors. -
Involvement in disease
Genetic variation in PDCD1 is associated with susceptibility to systemic lupus erythematosus type 2 (SLEB2) [MIM:605218]. Systemic lupus erythematosus is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system. -
Sequence similarities
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Developmental stage
Induced at programmed cell death. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 5133 Human
- Omim: 600244 Human
- SwissProt: Q15116 Human
- Unigene: 158297 Human
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Alternative names
- CD279 antibody
- CD279 antigen antibody
- hPD 1 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with ab228462 at 1/100 dilution (0.52 μg/ml) (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Panel A: merged staining of anti-Ki67 (magenta; Opal™690), anti-PD-L1 (red; Opal™570) and anti-PD1 (green; Opal™520) on human tonsil.
Panel B: anti-Ki67 stained on nucleus of proliferating cells.
Panel C: anti-PD-L1 stained on membrane of cells involved in T cell inhibition.
Panel D: anti-PD1 stained on antigen-stimulated T cells.
The section was incubated in three rounds of staining: in the order of ab16667 for 10 mins, ab243644 for 30 mins and ab228462 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] (ab243644)
Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human endometrial carcinoma stroma (PMID: 27446374) is observed.
The section was incubated with ab243644 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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All lanes : Anti-PD1 antibody [EPR23119-111] (ab243644) at 1/1000 dilution
Lane 1 : Untreated MOLT-4 (human lymphoblastic leukemia T lymphoblast)
Lane 2 : MOLT-4 treated with 500ng/ml Ionomycin and 10ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 30487606, 31783892).
Exposure time: 48 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), CD8 with ab178089 at 1/100 dilution (0.83 μg/ml) (C) and TIM 3 with ab242080 at 1/500 dilution (2.09 μg/ml) (B). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Panel A: merged staining of anti-TIM 3 (green; Opal™690), anti-CD8 (red; Opal™520) and anti-PD1 (gray; Opal™570) on human tonsil.
Panel B: anti-TIM 3 stained on membrane of a subset of immune cells.
Panel C: anti-CD8 stained on membrane of a subset of T cells.
Panel D: anti-PD1 stained on membrane of a subset of lymphocytes.
The section was incubated in three rounds of staining: in the order of ab242080, ab243644 and ab178089 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] (ab243644)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue labelling PDI with ab243644 at 1.02 µg/mL (B), PD-L 1 with ab213524 at 1/100 dilution (C) and CD68 with ab213363 at 1/300 dilution (D). Anti-Rabbit and Mouse Polymer HRP was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Heat mediated antigen retrieval (Leica ER2, PH9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibodies from the previous round, to avoid any cross-reactivity.
Panel A: merged staining of anti- PD1 (green, Opal™520), anti- PD-L1 (red, Opal™570) and anti- CD68 (yellow, Opal™690).
Panel B: Anti- PD1 stained on antigen-stimulated T cells.
Panel C: anti- PD-L1 stained on cells involved in T cell inhibition
Panel D: anti-CD68 stained on macrophages.
The section was incubated in three rounds of staining: in the order of ab243644, ab213363 and ab213524 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] (ab243644)
Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human Hodgkin's lymphoma (PMID: 16819321) is observed.
The section was incubated with ab243644 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] (ab243644)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on germinal center of human tonsil (PMID: 29042640, PMID: 16819321) is observed.
The section was incubated with ab243644 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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All lanes : Anti-PD1 antibody [EPR23119-111] (ab243644) at 1/1000 dilution
Lane 1 : Human tonsil tissue lysate
Lane 2 : Human tonsil tissue lysate treated with PNGase F
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 30487606, 31783892).
Exposure time: 48 seconds
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab243644 has been referenced in 2 publications.
- Rong X et al. PET/CT Imaging of Activated Cancer-Associated Fibroblasts Predict Response to PD-1 Blockade in Gastric Cancer Patients. Front Oncol 11:802257 (2021). PubMed: 35155199
- Zhu P et al. MiRNA505/NET1 Axis Acts as a CD8+ T-TIL Regulator in Non-Small Cell Lung Cancer. Onco Targets Ther 13:9785-9795 (2020). PubMed: 33061457