Search
Disclaimer
Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.
FIGURE: 1 / 3
CD19 Antibody (H004T03Y02-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD19 Monoclonal Antibody, NovaFluor Yellow 590 (right). All cells were co-stained with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0038-82). 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
H004T03Y02-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
HIB19
Conjugate
Excitation/Emission Max
563/594 nm
View spectra
Form
Liquid
Concentration
0.4 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 HIB19 monoclonal antibody reacts with human CD19, a 95 kDa transmembrane glycoprotein. The SJ25C1 antibody and the HIB19 monoclonal antibody recognize overlapping epitopes.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
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 (0.4 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
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.
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.
References (0)
Have you cited this product in a publication?
Let us know so we can reference it here.
Bioinformatics
Protein Aliases: B-lymphocyte antigen CD19; B-lymphocyte surface antigen B4; CD19; Differentiation antigen CD19; Leu-12; T-cell surface antigen Leu-12
Gene Aliases: B4; CD19; CVID3
UniProt ID: (Human) P15391
Entrez Gene ID: (Human) 930
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.*
Learn more
We're here to help
Get expert recommendations for common problems or connect directly with an on staff expert for technical assistance related to applications, equipment and general product use.
Contact tech support