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CD19 Antibody (M004T02Y02-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.2 µg of CD19 Monoclonal Antibody, NovaFluor Yellow 590 (right). All cells were co-stained with CD3e Monoclonal Antibody, eFluor 450 (Product # 48-0031-82). 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
M004T02Y02-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
eBio1D3 (1D3)
Conjugate
Excitation/Emission Max
563/594 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
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 eBio1D3 (1D3) monoclonal antibody reacts with mouse CD19, a 95 kDa transmembrane glycoprotein. CD19 is expressed by B cells during all stages of development excluding the terminally differentiated plasma cells. Follicular dendritic cells also express CD19. Together CD21, CD81, MHC class II, and CD19 form a multimolecular complex that associates with the BCR. Signaling through CD19 induces tyrosine phosphorylation, calcium flux and proliferation of B cells.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This eBio1D3 (1D3) antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.2 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 590 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: 563 nm; Emission: 594 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD19 is a member of the immunoglobulin superfamily, characterized by two Ig-like domains, and is expressed on B cells throughout all stages of development, excluding terminally differentiated plasma cells. It is also expressed on follicular dendritic cells and has been observed on myeloid leukemia cells, particularly those of monocytic lineage. CD19 is considered the earliest and broadest B cell-restricted antigen, and its expression is found in all B cell precursor leukemias. CD19 forms a multimolecular complex with CD21, CD81, Leu13, MHC class II, and the B cell receptor (BCR), playing a crucial role in B cell signaling. As a signal-amplifying coreceptor for the BCR, CD19 lowers the threshold for antigen receptor-dependent stimulation, allowing B cells to respond specifically and sensitively to various antigens through low-affinity antigen receptors. Signaling through CD19 induces tyrosine phosphorylation, calcium flux, and proliferation of B cells. Beyond its role as a BCR coreceptor, CD19 can also signal independently of BCR co-ligation, serving as a central regulatory component upon which multiple signaling pathways converge. This makes CD19 an important functional regulator of both normal and malignant B cell proliferation. Mutations in the CD19 gene can result in hypogammaglobulinemia, a condition characterized by low levels of immunoglobulins, while CD19 overexpression can lead to B cell hyperactivity. CD19 is expressed on 100% of peripheral B cells, as defined by the expression of kappa or lambda light chains, underscoring its significance in B cell function and immune regulation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: B-lymphocyte antigen CD19; CD19; Differentiation antigen CD19; Leu-12
Gene Aliases: AW495831; Cd19
UniProt ID: (Mouse) P25918
Entrez Gene ID: (Mouse) 12478
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