Anti-CD4 antibody [EPR6855] – Mouse IgG1 (Chimeric) – BSA and Azide Free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] – Mouse IgG1 (Chimeric) – BSA and Azide Free (AB317797)

This data was developed using the same antibody clone in a different buffer formulation (ab133616).

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4with ab133616 at a concentration of 1.43 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. ab133616 anti-CD4 antibody [EPR6855] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] – Mouse IgG1 (Chimeric) – BSA and Azide Free (AB317797)

Immunofluorescence staining of CD4 in sections of formalin-fixed paraffin-embedded human tonsil (positive) and human colon (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab317797 at 1/500 dilution, and then incubated for 1 hour with ab150112, Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] – Mouse IgG1 (Chimeric) – BSA and Azide Free (AB317797)

This data was developed using the same antibody clone in a different buffer formulation (ab133616).

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4with ab133616 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with OptiView DAB IHC Detection Kit followed by OptiView Amplification kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab133616 anti-CD4 antibody [EPR6855] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD4 antibody [EPR6855] – Mouse IgG1 (Chimeric) – BSA and Azide Free (AB317797)

ab317797 staining CD4 in THP-1 (positive) and HCT116 (negative) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab317797 at 1µg/ml. Cells were then incubated with ab150112, Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.