Description: The HIB19 monoclonal antibody reacts with human 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, Leu13, MHC class II, and CD19 form a multimolecular complex that associates with BCR. Signaling through CD19 induces tyrosine phosphorylation, calcium flux and proliferation of B cells. The SJ25C1 antibody and the HIB19 monoclonal antibody recognize overlapping epitopes.
Applications Reported: This HIB19 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This HIB19 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (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.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CD19 is a member of the immunoglobulin superfamily and has two Ig like domains. The CD19 molecule is expressed on 100% of the peripheral B cells as defined by expression of kappa or lambda light chains. CD19 appears to be expressed on myeloid leukemia cells, particularly those of monocytic lineage. Leukemia phenotype studies have demonstrated that the earliest and broadest B cell restricted antigen is the CD19 antigen. The receptor for CD19 is an important functional regulator of normal and malignant B cell proliferation, and is expressed in all B cell precursor leukemias. Lymphocytes proliferate and differentiate in response to various concentrations of different antigens. The ability of the B cell to respond in a specific, yet sensitive manner to the various antigens is achieved with the use of low-affinity antigen receptors. CD19 is a cell surface molecule which assembles with the antigen receptor of B lymphocytes in order to decrease the threshold for antigen receptor-dependent stimulation. Besides being a signal-amplifying coreceptor for the B cell receptor (BCR), CD19 can also signal independently of BCR co-ligation and is a central regulatory component upon which multiple signaling pathways converge. Mutation of the CD19 gene results in hypogammaglobulinemia, whereas CD19 overexpression causes B cell hyperactivity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: B-lymphocyte antigen CD19; B-lymphocyte surface antigen B4; CD19; Differentiation antigen CD19; T-cell surface antigen Leu-12
Gene Aliases: B4; CD19; CVID3
UniProt ID: (Human) P15391
Entrez Gene ID: (Human) 930