MAB3574
Anti-Vinculin Antibody, clone VIIF9 (7F9)
clone VIIF9 (7F9), Chemicon®, from mouse
Synonym(s):
Anti-CMD1W, Anti-CMH15, Anti-HEL114, Anti-MV, Anti-MVCL
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
purified antibody
antibody product type
primary antibodies
clone
VIIF9 (7F9), monoclonal
species reactivity
mouse, human, pig, rabbit, monkey, bovine
packaging
antibody small pack of 25 μg
manufacturer/tradename
Chemicon®
technique(s)
immunocytochemistry: suitable
immunohistochemistry: suitable (paraffin)
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG1
NCBI accession no.
UniProt accession no.
shipped in
ambient
target post-translational modification
unmodified
Gene Information
human ... VCL(7414)
Specificity
Vinculin. Cellular localization is Intracellular, associated with cytoskeleton.
Immunogen
Crude smooth muscle extract from normal human adult uterus.
Application
Anti-Vinculin Antibody, clone VIIF9 (7F9) is an antibody against Vinculin for use in IP, WB, IC, IH(P).
Research Category
Signaling
Signaling
Research Sub Category
Cytoskeletal Signaling
Cytoskeletal Signaling
Western blot. 1:1000; ~117kDa band. Suggested blocking buffer is TBS-Tween with 2% BSA. Suggested dilution buffer is TBS-Tween with 0.05% sodium azide. Preferred gel percentage is 10% and/or 4-20% (or similar) gradient gel.
Immunohistochemistry on frozen and paraffin embedded tissue sections. Suggested fixation for frozen tissue sections is acetone fix for 6 minutes at room temperature. For formalin fixed paraffin embedded tissue sections: microwave in 0.01M citrate buffer (pH 6.0) for 8-10 minutes (note that all microwaves differ and adjustments may need to be made). If necessary follow with enzyme digestion (0.01% pronase for 10 mintues). Suggested blocking agent is fetal bovine serum. The antibody has also been used successfully on methyl-Carnoy fixed tissue.
Immunocytochemistry (see application notes below).
Immunoprecipitation. Suggested extraction buffer is 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholic acid-NaCl and 0.5 mM PMSF. Final reaction volume is 1 mL and suggested capture agent is agarose conjugated anti-mouse IgG. Note that the muscle-specific isoform, meta-vinculin (MW 150 kDa) will co-precipitate.
Optimal working dilutions must be determined by the end user.
APPLICATION NOTES FOR MAB3574
IMMUNOCYTOCHEMISTRY
For cultured cells, use the following method: 1) wash cells twice with buffer "A" (100 mM Pipes (to make it dissolve, titrate with KOH), 0.5mM MgCl2, 0.1 mM EDTA, 0.01M EGTA (to make it dissolve, titrate with KOH), pH 6.9); 2) fix 15 min @RT (3.7% formalin, 0.2% Triton X-100, 2M Glycerol, prepared on buffer A); 3) wash 3 times with buffer "A"; 4) proceed with incubation with primary Abs, etc.
Immunohistochemistry on frozen and paraffin embedded tissue sections. Suggested fixation for frozen tissue sections is acetone fix for 6 minutes at room temperature. For formalin fixed paraffin embedded tissue sections: microwave in 0.01M citrate buffer (pH 6.0) for 8-10 minutes (note that all microwaves differ and adjustments may need to be made). If necessary follow with enzyme digestion (0.01% pronase for 10 mintues). Suggested blocking agent is fetal bovine serum. The antibody has also been used successfully on methyl-Carnoy fixed tissue.
Immunocytochemistry (see application notes below).
Immunoprecipitation. Suggested extraction buffer is 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholic acid-NaCl and 0.5 mM PMSF. Final reaction volume is 1 mL and suggested capture agent is agarose conjugated anti-mouse IgG. Note that the muscle-specific isoform, meta-vinculin (MW 150 kDa) will co-precipitate.
Optimal working dilutions must be determined by the end user.
APPLICATION NOTES FOR MAB3574
IMMUNOCYTOCHEMISTRY
For cultured cells, use the following method: 1) wash cells twice with buffer "A" (100 mM Pipes (to make it dissolve, titrate with KOH), 0.5mM MgCl2, 0.1 mM EDTA, 0.01M EGTA (to make it dissolve, titrate with KOH), pH 6.9); 2) fix 15 min @RT (3.7% formalin, 0.2% Triton X-100, 2M Glycerol, prepared on buffer A); 3) wash 3 times with buffer "A"; 4) proceed with incubation with primary Abs, etc.
Quality
Tested
Physical form
Format: Purified
Liquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide.
Storage and Stability
Maintain at 2-8°C in undiluted aliquots up to 6 months after date of receipt
Analysis Note
Control
POSITIVE CONTROL:
Vinculin is present in all cell types.
POSITIVE CONTROL:
Vinculin is present in all cell types.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Product No.
Description
Pricing
Storage Class Code
10 - Combustible liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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