Anti-F4/80 antibody [EPR26545-166] – Chicken IgY (Chimeric) – BSA and Azide Free

• IHC

Lab

Immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] – Chicken IgY (Chimeric) – BSA and Azide Free (AB320061)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab300421).

Immunohistochemical analysis of formalin fixed paraffin embedded mouse liver labelling F4/80 with ab300421 at a concentration of 0.03 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with anti-rabbit HQ and anti-HQ HRP followed by a ChromoMap DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.

ab300421 Anti-F4/80 antibody [EPR26545-166] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-F4/80 antibody [EPR26545-166] – Chicken IgY (Chimeric) – BSA and Azide Free (AB320061)

ab320061 staining F4/80 in Raw264.7 (positive) and NIH/3T3 (negative) cells. The cells were fixed with 100% methanol (5 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 ab320061 at 5µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-F4/80 antibody [EPR26545-166] – Chicken IgY (Chimeric) – BSA and Azide Free (AB320061)

Immunofluorescence staining of F4/80 in sections of formalin-fixed paraffin-embedded mouse spleen (positive) and mouse pancreas (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 ab320061 at 1/100 dilution and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). 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.