Immune Checkpoint Blockade: T Cell Responses

There is great interest in identifying downstream T cell responses following immune checkpoint blockade to direct more effective treatments and better-informed therapeutic decisions. R&D Systems and Tocris offer a wide selection of reagents, kits, and assays that will allow you to explore, modulate, and validate T cell responses downstream of immune checkpoint blockade.

Immune Checkpoint Blockade Homepage

 

Explore

Proteome Profiler™ Antibody Arrays

Analyze the expression levels or phosphorylation status of up to 111 proteins simultaneously. Imagine the time you’ll save investigating how your checkpoint blockade affects downstream responses!

Cytokine Profile of TNF-alpha-treated Balb/3T3 Cells. Balb/3T3 mouse embryonic fibroblast cells were untreated or treated with 100 ng/mL of Recombinant Mouse TNF-alpha (Catalog # 410-MT) for 24 hours. Cell supernates were analyzed using the Mouse XL Cytokine Array Kit (Catalog # ARY028).


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Signal Transduction Profile of IGF-I-treated MCF-7 Cells.  MCF-7 human breast cancer cells were either left untreated or treated with 100 ng/mL of Recombinant Human IGF-I (Catalog # 291-G1) for 1 hour. Cell lysates were analyzed using the Human Phospho-Kinase Array Kit (Catalog # ARY003B).

Luminex® Screening Assays

Utilize our Luminex® Screening Assays to analyze the expression levels of up to 100 analytes simultaneously! Our Screening Assays are cost-effective, accurate, flexible, and require only a small sample volume (‹50 µL).

 

Luminex Screening Assays Luminex Assay Ordering Tool

 

Modulate

Tocris provides high performance research tools that activate and inhibit key downstream mediators of T-cell signaling, allowing you to modulate and investigate key proteins involved in immune checkpoint blockade. Tocris small molecules and peptides are manufactured to industry leading standards of purity and quality.
Primary Target Cat. No. Name Description
Akt (Protein Kinase B) 4635 SC 79 Akt activator
AP-1 2476 SR 11302 AP-1 inhibitor
Brutons Tyrosine Kinase (BTK) 5012 PCI 29732 Potent BTK inhibitor
c-FMS (CSF1R) 4481 Ki 20227 c-Fms inhibitor
ERK 3706 FR 180204 Selective ERK inhibitor
G Proteins (Small) 4266 ML 141 Selective inhibitor of Cdc42 Rho family GTPase
Interleukin-2 inducible T cell kinase (ITK) 4710 PF 06465469 Potent inhibitor of ITK
IκB Kinase 4806 BMS 345541 Selective allosteric inhibitor of IKK; anti-inflammatory
MEK 4192 PD 0325901 Potent inhibitor of MEK1/2
mTOR 5264 Temsirolimus mTOR inhibitor
NFAT 3930 NFAT Inhibitor Inhibitor of calcineurin-mediated NFAT activation
p38 1402 SB 203580 hydrochloride Selective inhibitor of p38 MAPK
PI 3-Kinase 4674 CZC 24832 Selective inhibitor of PI 3-kinase γ
PP1 and PP2A 1336 Calyculin A Protein phosphatase 1 and 2A inhibitor
Src Family Kinases 5413 WH-4-023 Potent and selective Lck and Src inhibitor; also inhibits SIK
Syk Kinase 2471 ER 27319 maleate Selective Syk kinase inhibitor

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Validate

ELISAs

Trust the most-referenced ELISAs on the market to verify your antibody array and/or Luminex assay results. R&D Systems offers a wide variety of complete ELISA kits and ELISA Development Systems that have been exhaustively tested for superior quality and reproducibility.

Linearity of the Human IFN-gamma Quantikine® ELISA Kit (Catalog # DIF50). To assess the linearity of the assay, five samples were spiked with high concentrations of IFN-gamma in various matrices and diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

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Antibodies

Verify antibody array and Luminex assay results and generate publication-quality data with antibodies from R&D Systems. Our antibodies are manufactured and carefully tested with quality and performance as the top priorities. Learn more about what makes our antibodies the best.

Detection of IFN-gamma in Human PBMCs. Human PBMCs were stimulated with PMA (Catalog # 1201) and ionomycin. IFN-gamma was detected using Human IFN-gamma Monoclonal Antibody (Catalog # MAB285). Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Detection of TNF-alpha in Mouse T Cells. TNF-alpha was detected in immersion fixed activated mouse T cells using Mouse TNF-alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-410-NA). Cells were stained (red) and counterstained (green). Detection of Phospho-p38 by Western blot.  Lysates of HepG2 human hepatocellular carcinoma cells were untreated (-) or treated (+) with 20 ng/mL Recombinant Human IL-1 beta (Catalog # 201-LB) for the indicated times. The PVDF membrane was probed with Rabbit Anti-Human/Mouse/Rat Phospho-p38 MAP Kinase (T180/Y182) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF869), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008).

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ELISpot Kits and Development Modules

Identify and measure the frequency of cytokine secreting cells with the most accurate and sensitive ELISpot kits available. Our single and dual-color complete ELISpot kits are ready-to-run out of the box and require no assay development. Alternatively, we offer ELISpot Development Modules that contain the basic components required to develop your own ELISpot assay.

Identification of IFN-gamma- and IL-17-Secreting Cells. IFN-gamma (blue spots) and IL-17 (red spots) were secreted from mouse splenocytes stimulated with PMA/Ca2+ ionomycin. Spots of cytokine secretion were visualized using the Mouse IFN-gamma/IL-17 Dual-Color ELISpot Kit (Catalog # ELD5007).

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