Anti-AVPR1B 抗体 (ab104365)

All lanes : Anti-AVPR1B antibody (ab104365) at 1 µg/ml

Lane 1 : E10 Mouse Embryo Brain Tissue Lysate
Lane 2 : Forebrain (Mouse) Tissue Lysate
Lane 3 : Human brain normal tissue lysate - membrane extract (ab29456)
Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 5 : E10 Mouse Embryo Brain Tissue Lysate with Immunising peptide at 1 µg/ml
Lane 6 : Forebrain (Mouse) Tissue Lysate with Immunising peptide at 1 µg/ml
Lane 7 : Human brain normal tissue lysate - membrane extract (ab29456) with Immunising peptide at 1 µg/ml
Lane 8 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 46 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?
Additional bands at: 102 kDa, 22 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 30 seconds


VR1b contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.

ICC/IF image of ab104365 stained SKNSH 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 (ab104365, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) SKNSH cells at 1µg/ml.