CD28 (D2Z4E) & CO-0081-594 SignalStar™ Oligo-Antibody Pair #78824
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- IHC
SignalStar™ multiplex immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using MHC Class II (LGII-612.14) & CO-0105-488 SignalStar™ Oligo-Antibody Pair #55436 (green), E-Cadherin (24E10) & CO-0103-594 SignalStar™ Oligo-Antibody Pair #65400 (magenta), CD28 (D2Z4E) & CO-0081-647 SignalStar™ Oligo-Antibody Pair #96977 (red), α-Smooth Muscle Actin (D4K9N) & CO-0024-488 SignalStar™ Oligo-Antibody Pair #76133 (green), TCF1/TCF7 (C63D9) & CO-0006-594 SignalStar™ Oligo-Antibody Pair #34706 (yellow), and Arginase-1 (D4E3M) & CO-0075-647 SignalStar™ Oligo-Antibody Pair #44900 (white). All fluorophores have been assigned a pseudocolor, as indicated. Staining was performed on the BOND RX autostainer by Leica Biosystems.
Order Information # 78824
This product is not sold separately. Please see the SignalStar™ Multiplex IHC Panel Builder Tool for ordering information.
- Product Includes
- Related Products
| Product Includes | Volume | Reactivity | Isotype |
|---|---|---|---|
| CD28 (D2Z4E) Rabbit mAb (SignalStar™ Conjugate 0081) #50343 | 50 µl | H M | Rabbit IgG |
| Complementary Oligo (CO-0081-594) #65203 | 22 µl |
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| SignalStar™ Leica Bond | 1:50 - 1:200 |
| SignalStar™ Manual | 1:50 - 1:200 |
Storage
SignalStar conjugates are supplied in PBS (pH 7.2), less than 0.1% sodium azide, 2 mM EDTA, 0.05% Triton X-100, 2 mg/mL BSA, and 50% glycerol. Complementary oligos are supplied in nuclease-free water. Store at -20°C. Do not aliquot the antibody. All components in this kit are stable for at least 12 months when stored at the recommended temperature.
Product Description
SignalStar multiplex immunohistochemistry (IHC) is an advanced technology for labeling multiple proteins simultaneously in tissue samples using specific primary antibodies and fluorescent detection reagents. This technology offers accuracy and reliability in visualizing and analyzing protein expression while maintaining spatial context and tissue architecture.
SignalStar Oligo-Antibody Pairs are compatible with the SignalStar Multiplex IHC Buffer Kits for use in fluorescent multiplex imaging experiments. This product includes the oligo-conjugated antibodies and complementary oligos required for labeling your target protein on up to 10 slides. SignalStar Multiplex IHC Buffer Kits are required to amplify and image the target signal. Multiple oligo-antibody pairs can be conveniently combined into a multiplex panel using the SignalStar Multiplex IHC Panel Builder. SignalStar Multiplex IHC Kits & Reagents are not compatible with all of Cell Signaling Technology® products and protocols that are recommended for use in immunohistochemical assays.
SignalStar Oligo-Antibody Pairs are compatible with the SignalStar Multiplex IHC Buffer Kits for use in fluorescent multiplex imaging experiments. This product includes the oligo-conjugated antibodies and complementary oligos required for labeling your target protein on up to 10 slides. SignalStar Multiplex IHC Buffer Kits are required to amplify and image the target signal. Multiple oligo-antibody pairs can be conveniently combined into a multiplex panel using the SignalStar Multiplex IHC Panel Builder. SignalStar Multiplex IHC Kits & Reagents are not compatible with all of Cell Signaling Technology® products and protocols that are recommended for use in immunohistochemical assays.
Protocol
Specificity / Sensitivity
CD28 (D2Z4E) Rabbit mAb (SignalStar™ Conjugate 0081) recognizes endogenous levels of total CD28 protein and is predicted to react with multiple CD28 isoforms that are produced through alternative splicing. This antibody cross-reacts with 16 kDa and 100 kDa proteins of unknown origin in some mouse cells. Non-specific staining was observed in mouse testis by immunohistochemistry.
Species Reactivity:
Human, Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro208 of human CD28 protein.
Background
CD28 is a transmembrane glycoprotein expressed by T cells as well as some other hematopoietic cells (1,2). T cell activation requires T cell receptor (TCR) recognition of antigen presented in the context of MHC molecules. CD28 acts as a T cell costimulatory receptor, and interaction of CD28 with its ligands CD80 or CD86 provides the second signal required for naïve T cell activation (3-5). Activation of naïve T cells in the absence of CD28 stimulation can result in a state of T cell anergy, or unresponsiveness (3). CD28 signals through cytoplasmic phospho-tyrosine motifs that bind several SH2 or SH3 domain-containing proteins involved in T cell activation (2). Recently, CD28 was demonstrated to be a preferred target of PD-1-mediated dephosphorylation. Consistently, CD28 expression was required for T cell proliferation following PD-1 blockade and CD28 stimulation was required for effective anti-PD-1 cancer immunotherapy in mice (6,7). Several CD28 isoforms are produced by alternative splicing (8).
- Aruffo, A. and Seed, B. (1987) Proc Natl Acad Sci U S A 84, 8573-7.
- Esensten, J.H. et al. (2016) Immunity 44, 973-88.
- Harding, F.A. et al. (1992) Nature 356, 607-9.
- Azuma, M. et al. (1993) Nature 366, 76-9.
- Linsley, P.S. et al. (1990) Proc Natl Acad Sci U S A 87, 5031-5.
- Hui, E. et al. (2017) Science 355, 1428-1433.
- Kamphorst, A.O. et al. (2017) Science 355, 1423-1427.
- Magistrelli, G. et al. (1999) Biochem Biophys Res Commun 259, 34-7.
Pathways
Explore pathways related to this product.
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