Description: The RM4-5 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 RM4-5 is blocked by GK1.5.
Applications Reported: The RM4-5 antibody has been reported for use in flow cytometric analysis.
Applications Tested: The RM4-5 antibody has been tested by flow cytometric analysis of mouse thymocytes and splenocytes. This can be used at less than or equal to 0.5 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.
Super Bright 780 can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter, or equivalent. 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.
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.
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