Recombinant Anti-PD1 antibody [CAL20] (ab237728)
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-383701-ab237728pdl1pd1multiplexihcifantibodypanelhumantonsilimmunohistochemistry.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAL20] to PD1
- Suitable for: IHC-P, mIHC, WB, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PD1 antibody [CAL20]
See all PD1 primary antibodies -
Description
Rabbit monoclonal [CAL20] to PD1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, mIHC, WB, ICC/IFmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MOLT-4 (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h) whole cell lysate. ICC/IF: MOLT-4 cells (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h). IHC-P: Human tonsil, lymph node and lung carcinoma tissues. mIHC: Human tonsil tissue.
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: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Purification notes
Purity >99% -
Clonality
Monoclonal -
Clone number
CAL20 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab237728 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 |
|---|---|---|
| IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| mIHC |
Use at an assay dependent concentration.
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| WB |
1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 32 kDa).
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| ICC/IF |
1/50.
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| Notes |
|---|
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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mIHC
Use at an assay dependent concentration. |
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WB
1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 32 kDa). |
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ICC/IF
1/50. |
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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Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)
Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-345670-anti-pd1-antibody-cal20-immunohistochemistry-paraffin-tonsil-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)Immunohistochemical analysis of paraffin-embeded human tonsil tissue labeling PD1 with ab237728 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human tonsil.is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.
The section was incubated with ab237728 for 15 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-345671-anti-pd1-antibody-cal20-immunocytochemisrty-immunofluorescence-molt4-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] (ab237728)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MOLT-4 (human lymphoblastic leukemia cell line) cells labeling PD1 with ab237728 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in MOLT-4 cells treated with PMA (10ng/ml 24h) and Ionomycin(500ng/ml 24h). The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Cells were either untreated, ot treated with treated with PMA (10ng/ml 24h) and Ionomycin (500ng/ml 24h).
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![Western blot - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-345669-anti-pd1-antibody-cal20-western-blot-molt4-human.jpg)
Western blot - Anti-PD1 antibody [CAL20] (ab237728)All lanes : Anti-PD1 antibody [CAL20] (ab237728) at 1/1000 dilution
Lane 1 : Untreated MOLT-4 (human lymphoblastic leukemia cell line), whole cell lysate
Lane 2 : MOLT-4 (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 32 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-315298-anti-pd1-antibody-cal20-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)Formalin-fixed, paraffin-embedded human lymph node tissue stained for PD1 using ab237728 at 0.125 µg/ml dilution in immunohistochemical analysis.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-315299-anti-pd1-antibody-cal20-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)Formalin-fixed, paraffin-embedded human lung carcinoma tissue stained for PD1 using ab237728 at 0.125 µg/ml dilution in immunohistochemical analysis.
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![Anti-PD1 antibody [CAL20] (ab237728)](https://www.abcam.com/ps/products/237/ab237728/Images/ab237728-7-benefits-of-recombinant-antibodies.png)
Anti-PD1 antibody [CAL20] (ab237728)
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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Datasheet download
References (1)
ab237728 has been referenced in 1 publication.
- Chen Y et al. The frequency and inter-relationship of PD-L1 expression and tumour mutational burden across multiple types of advanced solid tumours in China. Exp Hematol Oncol 9:17 (2020). PubMed: 32775040
