PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812) is part of the multiplex kits range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
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Key facts
- Reactive species
Human
- Target
CD274
(See target data below)
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application mIHC | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813417, PubMed:28813410). As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response (PubMed:11015443, PubMed:28813417, PubMed:28813410). Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077).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:28813417, PubMed:28813410). The interaction with PDCD1/PD-1 inhibits cytotoxic T lymphocytes (CTLs) effector function (By similarity). 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 (By similarity).
Targets
PDCD1, CD68, CD3 epsilon, MKI67, KRT76
Alternative names
Programmed cell death 1 ligand 1, PD-L1, PDCD1 ligand 1, Programmed death ligand 1, hPD-L1, B7 homolog 1, B7-H1, B7H1, PDL1, PDCD1LG1, PDCD1L1, CD274
What's included?
Recommended products
PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812) is part of the multiplex kits range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
Key facts
- Reactive species
Human
- Target
CD274
- Applications
mIHC
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) is specifically designed and validated for multiplex analysis of the tumor microenvironment (TME) using tyramide signal amplification (TSA) on human formalin fixed paraffin-embedded tissue (FFPE) sections. The order of staining and antibody dilutions have been optimized on human tonsil FFPE tissue.
This panel empowers researchers to explore the spatial interactions of these important immuno-oncology biomarkers, immune checkpoint proteins and immune cell-phenotyping markers within the tumor micro-environment. It allows for the 6 targets to be fluorescently detected on the same tissue section, plus a nuclear counterstain (DAPI).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®). Please refer to this kit's protocol for directions on how to perform the immunostaining.
The panel contains enough reagents for a minimum of 50 slides.
Panel contents, recommended order of staining and dilutions:
1. Recombinant PD1 antibody [CAL20] ab237728 (50 μl)
1/500 dilution; Opal™520
2. Recombinant PD-L1 antibody [CAL10] ab237726 (50 μl)
1/500 dilution; Opal™540
3. Recombinant CD68 antibody [SP251] ab192847 - Macrophage marker (50 μl)
1/300 dilution; Opal™570
4. CD3 antibody [SP7] ab16669 - T cell marker (50 μl)
1/300 dilution; Opal™620
5. Recombinant Ki67 antibody [SP6] ab16667 - Proliferation marker (50 μl)
1/200 dilution; Opal™650
6. Pan-cytokeratin antibody [C-11] ab7753 - Epithelial tissue marker (50 μg)
1/200 dilution; Opal™690
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Terms & Conditions.
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Multiplex immunohistochemistry - PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812)
Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-PD1 (Anti-PD1 antibody [CAL20] ab237728; orange; Opal™520) anti-PDL1 (Anti-PD-L1 antibody [CAL10] ab237726; green; Opal™540) anti-CD68 (Anti-CD68 antibody [SP251] ab192847; yellow; Opal™570) anti-CD3 epsilon (Anti-CD3 epsilon antibody [SP7] ab16669; red; Opal™620) anti-Ki67 (Anti-Ki67 antibody [SP6] ab16667; light blue; Opal™650) and anti-PanCK (Anti-pan Cytokeratin antibody [C-11] 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 Anti-PD1 antibody [CAL20] ab237728 (1/500 dilution), Anti-PD-L1 antibody [CAL10] ab237726 (1/500 dilution), Anti-CD68 antibody [SP251] ab192847 (1/300 dilution), Anti-CD3 epsilon antibody [SP7] ab16669 (1/300 dilution), Anti-Ki67 antibody [SP6] ab16667 (1/200 dilution) and Anti-pan Cytokeratin antibody [C-11] 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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