Synthetic peptide corresponding to a region between residues 300 and 350 of human COL4A3BP (NP_005704.1)
In Western Blot, this antibody gave a positive signal in HeLa and 293T whole cell lysates.
This antibody gave a positive result when used in the following formaldehyde/methanol fixed cell lines: DU145.
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ICC/IF image of ab72536 stained DU145. The cells were 4% formaldehyde 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 ab72536 at (5 µg/mL) overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti Rabbit (96899) IgG (H+L) used at a 1/250 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.
Western blot - COL4A3BP antibody (ab72536)
All lanes : Anti-COL4A3BP antibody (ab72536) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
Predicted band size: 71 kDa Observed band size: 71 kDa Additional bands at: 100 kDa, 28 kDa, 60 kDa, 70 kDa (possible isoform). We are unsure as to the identity of these extra bands.
Immunoprecipitation - COL4A3BP antibody (ab72536)
Detection of Human COL4A3BP by Immunoprecipitation from Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab72536 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection.