Human Predicted to work with:
Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Pig, Xenopus laevis, Chimpanzee, Zebrafish, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
DomainThe START domain recognizes ceramide and mediates the intermembrane transfer of ceramide. The PH domain targets the Golgi apparatus. The FFAT motif is required for interaction with VAPA and VAPB.
Post-translational modificationsPhosphorylation on Ser-132 decreases the affinity toward phosphatidylinositol 4-phosphate at Golgi membranes and reduces ceramide transfer activity.
Cellular localizationCytoplasm. Golgi apparatus. Endoplasmic reticulum. Preferentially localized to the Golgi apparatus.
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.
References for Anti-COL4A3BP antibody (ab72536)
This product has been referenced in:
Yamaji T et al. Role of Intracellular Lipid Logistics in the Preferential Usage of Very Long Chain-Ceramides in Glucosylceramide. Int J Mol Sci17:N/A (2016).
Read more (PubMed: 27775668) »
Burgett AW et al. Natural products reveal cancer cell dependence on oxysterol-binding proteins. Nat Chem Biol7:639-47 (2011).
Read more (PubMed: 21822274) »