Anti-VAMP1 antibody [RP23040244]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VAMP1 antibody [RP23040244] (AB308137)

Immunofluorescence analysis of PC12 cells labeling endogenous VAMP1 with ab308137 at 5 µg/mL, followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1 : 2000 (Green). Nuclei (blue) is stained using DAPI. Cytoskeletal F-actin (red) staining using Rhodamine Phalloidin (1 : 300). Panels a-d show representative undifferentiated cells that were stained for detection and localization of VAMP1 protein (green) with no signal. Panels e-h clearly demonstrate enhanced signal of VAMP1 in cell body and neuronal processes of NGF (200 ng/mL 5 days) differentiated PC12 cells. The images were captured at 60X magnification.

• WB

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Western blot - Anti-VAMP1 antibody [RP23040244] (AB308137)

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of PC-12 (Lane 1), PC-12 treated with NGF and KCl (NGF at 200 nM, KCl at 50 nM for 7 days) (Lane 2) and tissue extracts (30 µg lysate) of Mouse Brain (Lane 3) and Rat Brain (Lane 4). The blots were probed with Anti-VAMP1 Recombinant Rabbit Polyclonal Antibody (ab308137, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.4 µg/mL, 1 : 4000 dilution). A 16 kDa band corresponding to VAMP1 was observed specifically in NGF differentiated PC-12 and tissues tested. Known quantity of protein samples were electrophoresed using a 10% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

Lane 1:

PC-12 whole cell extracts without treatment at 30 µg

Lane 2:

PC-12 whole cell extracts treated with NGF and KCl (NGF at 200 nM, KCl at 50 nM for 7 days) at 30 µg

Lane 3:

Mouse Brain tissue lysate at 30 µg

Lane 4:

Rat Brain tissue lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

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