Immunohistochemical analysis of paraffin-embedded human appendix using CD8α (C8/144B) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human lymph node using CD8α (C8/144B) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using CD8α (C8/144B) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human spleen using CD8α (C8/144B) Mouse mAb.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For EDTA: Heat slides in a microwave submersed in 1X EDTA unmasking solution until boiling is initiated; follow with 15 min at a sub-boiling temperature (95°-98°C). No cooling is necessary.
|SignalStain® Boost IHC Detection Reagent (HRP, Mouse) #8125||SignalStain® Boost IHC Detection Reagent (AP, Mouse) #31926|
|SignalStain® DAB Substrate Kit #8059||SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713|
|SignalStain® Vivid Purple Peroxidase Substrate Kit #96632|
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2015
revised June 2020
Protocol Id: 664
CD8α (C8/144B) Mouse mAb (IHC Specific) recognizes endogenous levels of total CD8 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CD8 protein.
Cluster of Differentiation 8 (CD8) is a disulphide-linked heterodimer consisting of the unrelated α and β subunits. Each subunit is a glycoprotein composed of a single extracellular Ig-like domain, a polypeptide linker, a transmembrane part and a short cytoplasmic tail. On T cells, CD8 is the coreceptor for the T cell receptor (TCR), and these two distinct structures recognize the Antigen–Major Histocompatibility Complex (MHC). Specifically, the Ig-like domain of CD8α interacts with the α3-domain of the MHC class I molecule. CD8 ensures specificity of the TCR–antigen interaction, prolongs the contact between the T cell and the antigen presenting cell, and the α chain recruits the tyrosine kinase Lck, which is essential for T cell activation (1).
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