711822

antibody from Invitrogen Antibodies

Targeting: VAMP1 SYB1, VAMP-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711822

Provider

Invitrogen Antibodies

Product name

VAMP1 Recombinant Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 (Product # 711822, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 Novex®NuPAGE®10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, PC12 cells were fixed and permeabilized for detection of endogenous VAMP1 using Anti-VAMP1 Recombinant Rabbit Polyclonal Antibody (Product # 711822, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) is stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938) and cytoskeletal F-actin (red) staining using Rhodamine Phalloidin (Product # R415, 1:300) Panel a-d) shows representative undifferentiated cells that were stained for detection and localization of VAMP1 protein (green) with no signal. Panel e-h) clearly demonstrate enhanced signal of VAMP1 in cell body and neuronal processes of NGF (200 ng/mL 5days) differentiated PC12 cells. The images were captured at 60X magnification.