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MAB1678 Sigma-Aldrich

Anti-Filamin A Antibody, clone PM6/317

ascites fluid, clone PM6/317, Chemicon®

Synonym: Alpha-Filamin, Filamin I, Endothelial Actin-binding Protein, ABP-280, Nonmuscle Filamin

  •  eCl@ss 32160702

  •  NACRES NA.41

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Properties

Related Categories Alphabetical Index, Antibodies, FB-FJ, Primary Antibodies
clone   PM6/317, monoclonal
biological source   mouse
application(s)   immunofluorescence: suitable
  immunohistochemistry: suitable (paraffin)
  immunohistochemistry: suitable
  immunoprecipitation (IP): suitable
  western blot: suitable
species reactivity   rat, rat, rabbit, human, guinea pig, mouse, chicken, human, mouse
shipped in   wet ice
antibody form   ascites fluid
isotype   IgG1
Quality Level   100
antibody product type   primary antibodies
mfr. no.   Chemicon®
NCBI accession no.   NM_001110556.1
UniProt accession no.   P21333

Description

General description

Filamin is a structural protein that forms flexible cross-links between two actin filaments. Filamin is a homodimer of polypeptide chains each joined to the other at one end with an actin binding site ath the other. It is present in smooth muscle, fibroblasts, platelets and lymphocytes.

Specificity

Expected to cross-react with chicken, guinea pig and rabbit.

Recognizes unprocessed, full length Human filamin (actin-binding protein; 270-280 kDa) as well as the 190 kDa N-terminal calpain cleavage fragment of filamin (Aakhus, 1992). Following induction of apoptosis in U937 cells, the antibody recognizes 170, 150, and 120 kDa N-terminal cleavage fragments of the full-length form presumably resulting from cleavage by activated caspase-3 (Umeda, 2001).

Immunogen

Human platelet protein.

Application

Immunoblotting:
1:1000-1:4000. Because of the large size of the unprocessed forms of filamin, 4-7% PAGE gels and proteinase inhibitors are recommended.

Immunofluorescence:
1:50-1:200 dilution from a previous lot was used. Suitable for staining both frozen and paraffin embedded tissues (at lower dilutions). Microwave-citrate buffer antigen retrieval method recommended for paraffin sections.

Immunoprecipitation:
A previous lot was used on immunoprecipitation. Suggested lysis buffer is PBS with 0.5% triton X-100 with proteinase inhibitors (note for full length filamin include calpain inhibitors). 5 microliters of antibody for every 300-500 μL of cell lysate (200-500 μg/mL total protein is suggested. Incubation is 1 hour RT or overnight 4C; Protein A/G agarose beads or rabbit anti-mouse secondary capture antibody is recommended for best recovery. 4-8% acrylamide gels are recommended for full length filamin or the 190 kDa fragement visualization.

Optimal working dilutions must be determined by end user.

Research Category
Cell Structure

Research Sub Category
Cytoskeleton

This Anti-Filamin A Antibody, clone PM6/317 is validated for use in IF, IH, IH(P), IP, WB for the detection of Filamin A.

Target description

270-280 kDa & 190 kDa

Physical form

Mouse monoclonal ascites IgG1 in buffer containing 0.1% sodium azide.

Unpurified

Storage and Stability

Stable for 1 year at -20°C in undiluted aliquots from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

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.

Quality

Routinely evaluated by Western Blot on Jurkat lysates.

Western Blot Analysis:
1:500-1:4000 dilution of this lot detected Filamin A on 10 μg of Jurkat lysates. Because of the large size of the unprocessed forms of filamin, 4-7% PAGE gels and proteinase inhibitors are recommended.

Analysis Note

Control
Positive control tisse: skin, jurkat cell lysate.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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