WB
Western blot analysis of CD4 on different lysates with Rabbit anti-CD4 antibody at 1/2,000 dilution. Lane 1: THP-1 WT cell lysate Lane 2: THP-1 CD4 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 51 kDa Observed band size: 55 kDa Exposure time: 30 seconds; ECL: Ori Supersensitive 4-20% SDS-PAGE gel. ET1609-52 was shown to specifically react with CD4 in THP-1 WT cells. Weakened band was observed when THP-1 CD4 KD sample was tested. THP-1 WT and THP-1 CD4 KD samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (1/2,000) and Loading control antibody (Rabbit anti-GAPDH, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CD4 antibody at 1/800 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of THP-1 cells labeling CD4 with Rabbit anti-CD4 antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CD4 antibody at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of THP-1 cells labeling CD4. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IF-P
Species: Human, Site: Spleen, Sample: Paraffin-embedded section, Antibody concentration: 1/200.| Product Name | CD4 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human CD4 aa 196-416 / 458. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-PIHCWB |
| WB:1:1000-1:2000 IHC:1:400-1:800 ICC/IF:1:50-1:200 FC:1:500-1:1000 IF-P:1:200 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CD4 |
|---|---|
| Gene Synonyms | T4 IMD79 Leu-3 OKT4D CD4mut |
| Gene Full Name | CD4 molecule |
| Gene Summary | This gene encodes the CD4 membrane glycoprotein of T lymphocytes. The CD4 antigen acts as a coreceptor with the T-cell receptor on the T lymphocyte to recognize antigens displayed by an antigen presenting cell in the context of class II MHC molecules. The CD4 antigen is also a primary receptor for entry of the human immunodeficiency virus through interactions with the HIV Env gp120 subunit. This gene is expressed not only in T lymphocytes, but also in B cells, macrophages, granulocytes, as well as in various regions of the brain. The protein functions to initiate or augment the early phase of T-cell activation, and may function as an important mediator of indirect neuronal damage in infectious and immune-mediated diseases of the central nervous system. Multiple alternatively spliced transcript variants encoding different isoforms have been identified in this gene. [provided by RefSeq, May 2020] |
| Molecular Weight(MW) | 51kDa |
| Cellular Localization | Cell membrane. |
WB
Western blot analysis of CD4 on different lysates with Rabbit anti-CD4 antibody at 1/2,000 dilution. Lane 1: THP-1 WT cell lysate Lane 2: THP-1 CD4 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 51 kDa Observed band size: 55 kDa Exposure time: 30 seconds; ECL: Ori Supersensitive 4-20% SDS-PAGE gel. ET1609-52 was shown to specifically react with CD4 in THP-1 WT cells. Weakened band was observed when THP-1 CD4 KD sample was tested. THP-1 WT and THP-1 CD4 KD samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (1/2,000) and Loading control antibody (Rabbit anti-GAPDH, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CD4 antibody at 1/800 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of THP-1 cells labeling CD4 with Rabbit anti-CD4 antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CD4 antibody at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of THP-1 cells labeling CD4. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IF-P
Species: Human, Site: Spleen, Sample: Paraffin-embedded section, Antibody concentration: 1/200.| Application Notes | WB:1:1000-1:2000 IHC:1:400-1:800 ICC/IF:1:50-1:200 FC:1:500-1:1000 IF-P:1:200 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Storage Buffer | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
