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 - Anti-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.
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.
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