This antibody gave a positive signal in Human Liver and Ovary tissue lysates and the following whole cell lysates; T47D, MCF7, HEK293, and HepG2.
This antibody gave a positive result in IHC in the following FFPE tissue: Human liver cancer.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
IHC image of PGRMC1 staining in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab80941, 1µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-PGRMC1 antibody (ab80941)
All lanes : Anti-PGRMC1 antibody (ab80941) at 1 µg/ml
Lane 1 : Human liver tissue lysate - total protein (ab29889) Lane 2 :T47D whole cell lysate (ab14899) Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 6 : Human ovary tissue lysate - total protein (ab30222)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Membrane-associated progesterone receptor component 1 (PGRMC1) contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight (25 kDa) than predicted (21 kDa).
ICC/IF image of ab80941 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab80941, 1µg/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.