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 Superclonal™ Antibody

Antibody type

Other

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Rabbit and Bovine based on sequence homology. Recombinant rabbit Superclonal™ antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit Superclonal™ antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production. Note: Formerly called “Recombinant polyclonal antibody”, this product is now rebranded as “Recombinant Superclonal™ antibody”. The physical product and the performance remain unchanged.

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 Superclonal™ Antibody (Product # 711822, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) 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 Superclonal™ Antibody (Product # 711822, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (Heavy Chain) 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.