Rabbit Recombinant Monoclonal PD1 antibody. Carrier free. Suitable for WB, mIHC, ICC/IF, IHC-P and reacts with Human, Rhesus monkey samples.
Images
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img123485.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613)")
![Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunocytochemistry-immunofluorescence-img86609.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img10835.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img91243.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401808.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (AB251613)")
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
Liquid
- Clonality
Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| IP | Flow Cyt | WB | mIHC | ICC/IF | IHC-P | |
|---|---|---|---|---|---|---|
| Human | Not recommended | Not recommended | Tested | Tested | Tested | Tested |
| Rhesus monkey | Not recommended | Not recommended | Expected | Expected | Expected | Tested |
IP
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rhesus monkey | Dilution info - | Notes - |
Flow Cyt
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Rhesus monkey | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rhesus monkey | Dilution info Use at an assay dependent concentration. | Notes - |
mIHC
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rhesus monkey | Dilution info Use at an assay dependent concentration. | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rhesus monkey | Dilution info Use at an assay dependent concentration. | Notes - |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rhesus monkey | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Associated Products
Select an associated product type
4 products for Alternative Product
Target data
Function
Inhibitory receptor on antigen activated T-cells that plays a critical role in induction and maintenance of immune tolerance to self (PubMed:21276005). Delivers inhibitory signals upon binding to ligands CD274/PDCD1L1 and CD273/PDCD1LG2 (PubMed:21276005). Following T-cell receptor (TCR) engagement, PDCD1 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (By similarity). Suppresses T-cell activation through the recruitment of PTPN11/SHP-2: following ligand-binding, PDCD1 is phosphorylated within the ITSM motif, leading to the recruitment of the protein tyrosine phosphatase PTPN11/SHP-2 that mediates dephosphorylation of key TCR proximal signaling molecules, such as ZAP70, PRKCQ/PKCtheta and CD247/CD3zeta (By similarity).The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28951311). The interaction with CD274/PDCD1L1 inhibits cytotoxic T lymphocytes (CTLs) effector function (PubMed:28951311). The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (PubMed:22658127, PubMed:25034862, PubMed:25399552).
Alternative names
Programmed cell death protein 1, Protein PD-1, hPD-1, PD1, PDCD1
Recommended products
Rabbit Recombinant Monoclonal PD1 antibody. Carrier free. Suitable for WB, mIHC, ICC/IF, IHC-P and reacts with Human, Rhesus monkey samples.
Alternative names
Programmed cell death protein 1, Protein PD-1, hPD-1, PD1, PDCD1
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
Liquid
- Clonality
Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
Yes
- Clone number
CAL20
- Purification technique
Affinity purification Protein A
- Concentration
- Purification notes
Purity is greater than 99%.
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
- Storage information
Do Not Freeze
Notes
ab251613 is the carrier-free version of Anti-PD1 antibody [CAL20] ab237728.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
PD1 also known as Programmed Cell Death Protein 1 or PDCD1 is a transmembrane protein that plays a critical role in regulating immune responses. It has a mass of approximately 55 kDa. PD1 is expressed on the surface of T cells B cells and some myeloid cells. PD1’s expression increases upon activation of these immune cells assisting in maintaining peripheral tolerance. Researchers often use PD1 mouse models and chimeric antibodies to explore the function of PD1 for experimental purposes. Antibodies such as anti-PD1 such as EH12.2H7 help in blocking PD1 interaction to study its role further.
- Biological function summary
PD1 serves as an inhibitory receptor acting as a checkpoint in the immune system. It becomes part of an immune-suppressive complex when it binds with its ligands PD-L1 or PD-L2 which are expressed on various cell types including some tumor cells. This interaction suppresses the proliferation of T cells and cytokine production contributing to immune homeostasis. By controlling T cell activity PD1 limits autoimmunity but can also reduce the immune system's capability to attack cancer cells.
- Pathways
PD1 functions in the immune checkpoint pathway a critical regulatory circuit in immune regulation. The engagement of PD1 with its ligands initiates a cascade that inhibits the function and proliferation of T cells through downstream SHP-2 phosphatase activity. This pathway frequently involves other regulatory proteins like CTLA-4 and is an important mechanism by which the body modulates immune responses. Related pathways often intersect with those involving T cell receptor signaling and contribute to the overall modulation of immune activity.
- Associated diseases and disorders
PD1 has a significant role in cancer and autoimmune disorders. PD1 expression can allow tumors to evade immune surveillance making PD1 a target for cancer therapies such as anti-PD1 antibodies which aim to block PD1 and restore T cell activity. The interaction of PD1 with cancer-related proteins like PD-L1 facilitates tumor immune evasion. In autoimmune disorders PD1’s regulation of immune balance can become dysregulated leading to persistent immune activation and tissue damage. Understanding PD1 and its interaction with proteins such as PD-L1 helps in developing therapeutic strategies for both cancer and autoimmune conditions.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
12 product images
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img123485.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Immunohistochemical analysis of paraffin-embeded human tonsil tissue labeling PD1 with Anti-PD1 antibody [CAL20] 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 Anti-PD1 antibody [CAL20] ab237728 for 15 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
![Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunocytochemistry-immunofluorescence-img86609.jpg "Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MOLT-4 (human lymphoblastic leukemia cell line) cells labeling PD1 with Anti-PD1 antibody [CAL20] ab237728 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img10835.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Formalin-fixed, paraffin-embedded human lymph node tissue stained for PD1 using Anti-PD1 antibody [CAL20] ab237728 at 0.125 μg/ml dilution in immunohistochemical analysis.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img91243.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Formalin-fixed, paraffin-embedded human lung carcinoma tissue stained for PD1 using Anti-PD1 antibody [CAL20] ab237728 at 0.125 μg/ml dilution in immunohistochemical analysis.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401808.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of ImmunoAtlas.Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611; cyan; Opal™ 520), Anti-Granzyme B (Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485; red; Opal™ 620), Anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894; red; Opal™ 620), Anti-CD8 alpha (Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596; green; Opal™ 650) and Anti-FOXP3 (Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611 (1/750 dilution), Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803 (1/250 dilution), ab251613 (1/750 dilution), Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485 (0.5 μg/ml), Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894 (1/1250 dilution), Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596 (1/1500 dilution) and Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048 (10 μg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 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 3 Imaging System (Akoya Biosciences®).This data is courtesy of ImmunoAtlas and it can be found here.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img422384.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of Dr. Chi Ngai ChanImmunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Immunohistochemical analysis of paraffin-embeded Rhesus monkey tonsil tissue labeling PD1 with ab251613 followed by Polink 1 Polymer HRP anti-Rabbit IgG.
Heat mediated antigen retrieval-Buffer/Enzyme Used: Dako pH9.
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401812.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of ImmunoAtlas.Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611; cyan; Opal™ 520), Anti-Granzyme B (Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485; red; Opal™ 620), Anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894; red; Opal™ 620), Anti-CD8 alpha (Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596; green; Opal™ 650) and Anti-FOXP3 (Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611 (1/750 dilution), Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803 (1/250 dilution), ab251613 (1/750 dilution), Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485 (0.5 μg/ml), Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894 (1/1250 dilution), Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596 (1/1500 dilution) and Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048 (10 μg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 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 3 Imaging System (Akoya Biosciences®).This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
This data is courtesy of ImmunoAtlas and it can be found here.
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401809.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of ImmunoAtlas.Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611; cyan; Opal™ 520), Anti-Granzyme B (Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485; red; Opal™ 620), Anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894; red; Opal™ 620), Anti-CD8 alpha (Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596; green; Opal™ 650) and Anti-FOXP3 (Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611 (1/750 dilution), Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803 (1/250 dilution), ab251613 (1/750 dilution), Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485 (0.5 μg/ml), Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894 (1/1250 dilution), Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596 (1/1500 dilution) and Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048 (10 μg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 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 3 Imaging System (Akoya Biosciences®).This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
This data is courtesy of ImmunoAtlas and it can be found here.
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img423097.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Tissue Microarrays stained for "Anti-PD1 antibody [CAL20]" using "Anti-PD1 antibody [CAL20] ab237728"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes. The sections were incubated with Anti-PD1 antibody [CAL20] ab237728 for 15 mins at room temperature followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img435960.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue staining PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a 1:500 (2.19 ug/ml) dilution; PD1 with Anti-PD1 antibody [CAL20] ab237728 at 1:2000 (0.525 ug/ml) dilution and CD68 with Anti-CD68 antibody [EPR20545] ab213363 at 1:500 (1.26 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.
Panel A: merged staining of anti-CD68 (magenta; Opal™690), anti-PD-L1 (green; Opal™520) and anti-PD1 (red; Opal™570) on human tonsil.
Panel B: anti-PD-L1 stained on cells involved in T cell inhibition.
Panel C: anti-PD1 stained on antigen-stimulated T cells.
Panel D: anti-CD68 stained on macrophages.The section was incubated in three rounds of staining: in the order of Anti-CD68 antibody [EPR20545] ab213363 and Anti-PD-L1 antibody [28-8] ab205921 for 30 mins, then Anti-PD1 antibody [CAL20] ab237728 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.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401810.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of ImmunoAtlas.Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611; cyan; Opal™ 520), Anti-Granzyme B (Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485; red; Opal™ 620), Anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894; red; Opal™ 620), Anti-CD8 alpha (Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596; green; Opal™ 650) and Anti-FOXP3 (Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611 (1/750 dilution), Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803 (1/250 dilution), ab251613 (1/750 dilution), Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485 (0.5 μg/ml), Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894 (1/1250 dilution), Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596 (1/1500 dilution) and Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048 (10 μg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 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 3 Imaging System (Akoya Biosciences®).This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
This data is courtesy of ImmunoAtlas and it can be found here.
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401811.jpg "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
This image is courtesy of ImmunoAtlas.Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of Anti-PD-L1 (Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611; cyan; Opal™ 520), Anti-Granzyme B (Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485; red; Opal™ 620), Anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894; red; Opal™ 620), Anti-CD8 alpha (Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596; green; Opal™ 650) and Anti-FOXP3 (Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.
The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; sequentially for Anti-PD-L1 antibody [CAL10] - BSA and Azide free ab251611 (1/750 dilution), Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free ab219803 (1/250 dilution), ab251613 (1/750 dilution), Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free ab264485 (0.5 μg/ml), Anti-EpCAM antibody [EPR20532-225] - BSA and Azide free ab225894 (1/1250 dilution), Anti-CD8 alpha antibody [CAL66] - BSA and Azide free ab251596 (1/1500 dilution) and Anti-FOXP3 antibody [236A/E7] - BSA and Azide free ab96048 (10 μg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 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 3 Imaging System (Akoya Biosciences®).This data is courtesy of ImmunoAtlas and it can be found here.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-PD1 antibody [CAL20] ab237728).
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img422384.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401812.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401809.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img423097.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img435960.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401810.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
![Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613), expandable thumbnail](https://content.abcam.com/products/images/pd1-antibody-cal20-bsa-and-azide-free-ab251613--multiplex-immunohistochemistry-img401811.jpg/_jcr_content/renditions/webp-475.webp "Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)")
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