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CD4 Antibody (M001T02Y02-A) in Flow
NovaFluor Yellow 590 non-specific staining of B cells in the SJL strain of mice. Splenocytes from C57BL/6 (left) and SJL (right) strains of mice were stained with Anti-Mouse CD45R (B220) Monoclonal Antibody conjugated to eFluor 450 and a non-cross-reactive, human-specific monoclonal antibody conjugated to NovaFluor Yellow 590. These data show that NovaFluor Yellow 590-conjugated antibodies non-specifically stain B cells in SJL mice (right). Non-specific staining has not been observed in C57BL/6 (left) mice and BALB/c mice (data not shown).
Product Details
M001T02Y02-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
GK1.5
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
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
Description
The GK1.5 monoclonal antibody reacts with the mouse CD4 molecule, a 55 kDa cell surface receptor expressed by a majority of thymocytes, subpopulation of mature T cells and dendritic cells. CD4 binds to MHC class II on the surface of antigen presenting cells and plays an important role both in T cell development and in optimal functioning of mature T cells. In T cells, CD4 associates with protein tyrosine kinase p56lck through its cytoplasmic tail. Binding of GK1.5 is blocked by RM4-5.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This GK1.5 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can 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
The CD4 antigen is involved in the recognition of MHC class II molecules and is a co-receptor for HIV. CD4 is primarily expressed in a subset of T-lymphocytes, also referred to as T helper cells, but may also be expressed by other cells in the immune system, such as monocytes, macrophages, and dendritic cells. At the tissue level, CD4 expression may be detected in thymus, lymph nodes, tonsils, and spleen, and also in specific regions of the brain, gut, and other non-lymphoid tissues. CD4 functions to initiate or augment the early phase of T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase, Lck. It may also function as an important mediator of direct neuronal damage in infectious and immune-mediated diseases of the central nervous system. Multiple alternatively spliced transcripts have been identified in this gene [RefSeq, July 2017].
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: CD4; CD4 antigen p55; cd4a; fCD4; Leu-3; T-cell differentiation antigen L3T4; T-cell surface antigen T4/Leu-3; T-cell surface glycoprotein CD4; T-cell surface glycoprotein CD4 precursor (T-cell surface antigen T4/Leu-3) (T-cell differentiation antigen L3T4)
Gene Aliases: Cd4; L3T4; Ly-4
UniProt ID: (Mouse) P06332
Entrez Gene ID: (Mouse) 12504
cytokine production
immune system process
induction by virus of host cell-cell fusion
immune response
cell adhesion
cell surface receptor signaling pathway
T cell differentiation
maintenance of protein location in cell
T cell activation
T cell selection
protein palmitoleylation
positive regulation of protein kinase activity
positive regulation of peptidyl-tyrosine phosphorylation
defense response to Gram-negative bacterium
positive regulation of calcium-mediated signaling
regulation of T cell activation
positive regulation of T cell activation
Performance Guarantee
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