Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (AB150112)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (AB150112)

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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (AB150112)

HeLa cells showing negative staining by ICC/IF using only secondary antibody. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The secondary antibody (orange) was ab150112 Alexa Fluor® 594 donkey anti-mouse IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (AB150112)

ICC/IF image of ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7291, 1µg/ml) overnight at +4°C. The secondary antibody (orange) was ab150112 Alexa Fluor® 594 donkey anti-mouse IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• Alexa Fluor®

Unknown

Alexa Fluor® - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (AB150112)