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Invitrogen
TCR V alpha 2 Monoclonal Antibody (B20.1), NovaFluor™ Yellow 610, eBioscience™
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FIGURE: 1 / 3
TCR V alpha 2 Antibody (M033T03Y03-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.1 µg of TCR V alpha 2 Monoclonal Antibody, NovaFluor Yellow 610 (right). All cells were co-stained with CD4 Monoclonal Antibody, eFluor 450 (Product # 48-0041-82) and CD8a Monoclonal Antibody, eFluor 450 (Product # 48-0081-82). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
M033T03Y03-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2a, lambda
Class
Monoclonal
Type
Antibody
Clone
B20.1
Conjugate
Excitation/Emission Max
551/614 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
AB_3099098 (AB_2926223)
Product Specific Information
Description
The B20.1 monoclonal antibody reacts with mouse T-cell receptor (TCR) V alpha 2 chains. This antibody recognizes the majority of the TCR V alpha 2 subfamily in mice carrying the a, b and c haplotypes. The antibody also reacts with the products of T cell receptor, V delta 8 due to the high degree of homology.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This B20.1 antibody has been tested by flow cytometric analysis of mouse lymph node cells. This can be used at less than or equal to 0.1 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 to preblock Fc receptors for 10 minutes using 0.5 µg of purified anti-mouse CD16/32 (Product # 14-0161-82). It is also 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 (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 610 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: 614 nm; Laser: 561 nm (Yellow) Laser
Target Information
The ability of T cell receptors (TCR) to discriminate foreign from self-peptides presented by major histocompatibility complex (MHC) class II molecules is essential for an effective adaptive immune response. TCR recognition of self-peptides has been linked to autoimmune disease. Mutant self-peptides have been associated with tumors. Engagement of TCRs by a family of bacterial toxins know as superantigens has been responsible for toxic shock syndrome. Autoantibodies to V beta segments of T cell receptors have been isolated from patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The autoantibodies block TH1-mediated inflammatory autodestructive reactions and are believed to be a method by which the immune system compensates for disease. Most human T cells express the TCR alpha-beta and either CD4 or CD8 molecule (single positive, SP). A small number of T cells lack both CD4 and CD8 (double negative, DN). Increased percentages of alpha-beta DN T cells have been identified in some autoimmune and immunodeficiency disorders. Gamma-delta T cells are primarily found within the epithelium. They show less TCR diversity and recognize antigens differently than alpha-beta T cells. Subsets of gamma-delta T cells have shown antitumor and immunoregulatory activity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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