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Invitrogen
CD20 Monoclonal Antibody (2H7), NovaFluor™ Blue 725, eBioscience™
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FIGURE: 1 / 2
CD20 Antibody (H076T03B10-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD20 Monoclonal Antibody, NovaFluor Blue 725 (right). All cells were co-stained with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0038-42). 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
H076T03B10-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
2H7
Conjugate
Excitation/Emission Max
492/725 nm
View spectra
Form
Liquid
Concentration
0.6 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
Description
The 2H7 monoclonal antibody reacts with human CD20, a 33-36 kDa transmembrane protein. CD20 is expressed by developing B cells as well as mature B cells but not plasma cells. CD20 has been detected at low levels on a small subset of mature T cells. It is suggested that CD20 plays a role in B-cell activation.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The 2H7 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 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.
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
Excitation: 493 nm; Emission: 726 nm; Laser: 488 nm (Blue) Laser
Target Information
CD20 is a non-glycosylated surface phosphoprotein that has a molecular weight range of 33-37 kDa depending on the degree of phosphorylation. CD20 is expressed on mature and most malignant B cells, in a subpopulation of T lymphocytes and follicular dendritic cells. CD20 expression on B cells is synchronous with the expression of surface IgM and it regulates transmembrane calcium conductance, cell cycle progression and B-cell proliferation. CD20 is also associated with lipid rafts, but the intensity of this association depends on extracellular triggering, employing CD20 conformational change, and/or BCR (B cell antigen receptor) aggregation. After the receptor ligation, BCR and CD20 colocalize and then rapidly dissociate before BCR endocytosis, whereas CD20 remains at the cell surface. CD20 serves as a useful target for antibody-mediated therapeutic depletion of B cells, as it is expressed at high levels on most B-cell malignancies, but does not become internalized or shed from the plasma membrane following monoclonal antibody treatment. Diseases associated with CD20 dysfunction include Ms4a1-related common variable immune deficiency.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: APY; ATOPY; B-lymphocyte antigen CD20; B-lymphocyte cell-surface antigen B1; B-lymphocyte surface antigen B1; Bp35; CD20; CD20 antigen; CD20 receptor; Fc epsilon receptor I beta chain; Fc Fragment of IgE high affinity I receptor for beta polypeptide; FCER1B; High affinity immunoglobulin epsilon receptor subunit beta; IgE Fc receptor subunit beta; IGEL; IGER; IGHER; LEU16; Leukocyte surface antigen Leu-16; Ly44; Membrane-spanning 4-domains subfamily A member 1; membrane-spanning 4-domains, subfamily A, member 1; MGC3969
Gene Aliases: B1; Bp35; CD20; CVID5; LEU-16; MS4A1; MS4A2; S7
UniProt ID: (Human) P11836
Entrez Gene ID: (Human) 931
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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