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CD8a Antibody (M003T02Y07-A) in Flow
C57BL/6 mouse splenocytes were either left unstained (blue histogram) or stained with 0.5 µg of CD8a Monoclonal Antibody, NovaFluor Yellow 730 (purple histogram) and acquired in the B1 channel on a 5-laser Cytek Aurora. Cells in the lymphocyte gate were used in the analysis.
Product Details
M003T02Y07-A
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Published species
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
53-6.7
Conjugate
Excitation/Emission Max
551/730 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
Description
The 53-6.7 monoclonal antibody reacts with the mouse CD8a molecule.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 53-6.7 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 730 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 551 nm; Emission: 730 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD8, also known as cluster of differentiation 8, is a type I transmembrane glycoprotein of the immunoglobulin family that plays a crucial role in T cell differentiation, activation, and signal transduction. It is expressed as either a heterodimer (CD8 alpha beta) or a homodimer (CD8 alpha alpha). The CD8 alpha beta form is predominantly found on the majority of thymocytes and a subpopulation of mature alpha beta TCR T cells, while the CD8 alpha alpha form is expressed on gamma delta TCR T cells, a subset of intestinal intraepithelial lymphocytes (IELs), and dendritic cells. CD8 functions as a co-receptor for major histocompatibility complex class I (MHC-I) molecules, working alongside the T cell receptor (TCR). The CD8 alpha chain is essential for binding to MHC-I. CD8 is also expressed on a subset of T cells, NK cells, monocytes, and dendritic cells as disulfide-linked homodimers of CD8 alpha. Upon ligation of MHC-I/peptide complexes presented by antigen-presenting cells (APCs), CD8 recruits lymphocyte-specific protein tyrosine kinase (Lck), leading to lymphokine production, increased motility, and activation of cytotoxic T lymphocytes (CTLs). Activated CTLs are vital for clearing pathogens and tumor cells. The differentiation of naive CD8+ T cells into CTLs is strongly enhanced by cytokines such as IL-2, IL-12, and TGF-beta1. Through its interactions with MHC-I and association with protein tyrosine kinase p56lck, CD8 plays a significant role in T cell development and the activation of mature T cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: CD8a; CD8alpha; CD8b; CD8beta; fCD8; Leu-2; leu-2a; Lymphocyte antigen 3; Lyt-2.1 lymphocyte differentiation antigen (AA at 100); T-cell membrane glycoprotein Ly-3; T-cell surface glycoprotein CD8 alpha chain; T-cell surface glycoprotein CD8 beta chain; T-cell surface glycoprotein Lyt-2; T-cell surface glycoprotein Lyt-3
Gene Aliases: BB154331; Cd8a; Cd8b; Cd8b1; Ly-2; Ly-3; Ly-35; Ly-B; Ly-C; Lyt-2; Lyt-3; Lyt2; Lyt3
UniProt ID: (Mouse) P01731, (Mouse) P10300
Entrez Gene ID: (Mouse) 12525, (Mouse) 12526
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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