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MAB3574

Sigma-Aldrich

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

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)

Related Categories

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
Research Sub Category
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.

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.

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.

wgk_germany

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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