Anti-C4d抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4d antibody (AB87424)

IHC image of C4d staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab87424, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-C4d antibody (AB87424)

ICC image of ab87424 stained HepG2 cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat 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 (ab87424, 5μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM. This antibody also gave a positive result in 100% Methanol fixed (5 min) HeLa, and MCF-7 cells at 5μg/ml.

• WB

Unknown

Western blot - Anti-C4d antibody (AB87424)

C4d circulates in blood as a disulfide-linked trimer of an alpha, beta and gamma chain. Our immunogen sequence is within the alpha chain. The band at 85kDa could correspond to the alpha chain.

All lanes:

Western blot - Anti-C4d antibody (ab87424) at 1 µg/mL

All lanes:

Human Plasma Total Protein Lysate (ab83997) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

Predicted band size: 193 kDa

Observed band size: 110 kDa,130 kDa,193 kDa,30 kDa,48 kDa,85 kDa

true

Exposure time: 30s