CD4 Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of CD4 using anti-CD4 antibody (MBS1753316).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: mouse RAW264. 7 whole cell lysates
Lane 2: mouse ANA-1 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-CD4 antigen affinity purified polyclonal antibody (Catalog # MBS1753316) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for CD4 at approximately 52KD. The expected band size for CD4 is at 52KD.

Figure 2. IHC analysis of CD4 using anti-CD4 antibody (MBS1753316).
CD4 was detected in paraffin-embedded section of mouse thymus tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD4 Antibody (MBS1753316) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen.

Figure 3. Flow Cytometry analysis of mouse PBMC cells using anti-CD4 antibody (MBS1753316).
Overlay histogram showing mouse PBMC cells stained with MBS1753316 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD4 Antibody (MBS1753316, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.