Western blot analysis of CD4 was performed by loading 25 µg of Jurkat (lane 1) and CTLL-2 (lane 2) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a CD4 monoclonal antibody (Product # MA5-12259) at a dilution of 1:20 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~55 kDa.
Formalin-fixed, paraffin-embedded human tonsil stained with CD4 antibody using peroxidase-conjugate and DAB chromogen. Note membrane staining of T cells.
Flow cytometry analysis of CD4 in K562 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CD4 monoclonal antibody (Product # MA5-12259) at a dilution of 1:2 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
Flow cytometry analysis of CD4 in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-12259) at a dilution of 1:2. Cells were incubated for 30 min at 4°C and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4°C in the dark. FACS analysis was performed using 400 µL of cell buffer.
Flow cytometry analysis of CD4 in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-12259) at a dilution of 1:2. Cells were incubated for 30 min at 4°C and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4°C in the dark. FACS analysis was performed using 400 µL of cell buffer.
MA5-12259 targets CD4 in IHC (P), WB and FACS applications and shows reactivity with Human samples.
The MA5-12259 immunogen is stimulated human leukocytes.
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].
蛋白别名: CD4; CD4 antigen (p55); CD4 antigen p55; CD4 receptor; cd4a; fCD4; Leu-3; T-cell surface antigen T4/Leu-3; T-cell surface glycoprotein CD4
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