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MAB3568

Sigma-Aldrich

Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone ID8

clone ID8 (SMMS-1), Chemicon®, from mouse

Synonym(s):
Anti-CCDC128, Anti-KLRAQ1
eCl@ss:
32160702
NACRES:
NA.41

Quality Level

biological source

mouse

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

ID8 (SMMS-1), monoclonal

species reactivity

human, bovine, pig, rabbit

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

wet ice

Gene Information

human ... MYH11(4629)

Application

Western blot (see application notes below). Suggested blocking buffer is TBS-Tween with 2% BSA. Suggested dilution buffer is TBS-Tween with 0.05% sodium azide. Preferred gel percentage is 5% (see application notes).

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) follow with enzyme digestion (0.01% pronase for 10 mintues). Suggested blocking agent is fetal bovine serum.

Immunocytochemistry

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.

Optimal working dilutions must be determined by the end user.

APPLICATION NOTES FOR MAB3568

WESTERN BLOT

To achieve good resolution of myosin heavy chain isoforms with distinct molecular weight (200 - 2004 kDa), the following procedure should be followed: 1). Pyrophoshate extraction buffer for sample preparation (see below); run SDS-PAGE in 5% gel. Important: for better resolution of the MHC bands, use electrophoretic buffer with pH 8.2 (i.e. 0.1 less than standard), and prepare resolving gel (5%) with pH 9.0 (not 8.8 as usual). Also help thorough degasing of the resolving gel mixture (H2O, acrylamide, EDTA, pH 9.0, before (!) adding SDS, TEMED and APS). Run SDS-PAGE longer than after the dye front runs off (use 200 kDa MW markers and let it′s 200 kDa band run at least to the middle of 8X8 gel (using a big size gel (not the mini-gel!) will enhance the quality of MHC band resolution).

Pyrophosphate extraction buffer: (40mM Na4P2O7x10H2O, 1mM MgCl2, 1mM EGTA (add KOH to dissolve EGTA), PMSF, pH 9.5). To extract acto-myosin from tissues/cells, shake minced tissue or cells in cold extraction buffer 1 hr on ice bath (0oC), centrifuge @10,000g for 10 min at +2-8 oC, take supernatant and mix it 1:1 with standard Laemmli sample buffer, boil, run SDS-PAGE in 5% gel (see above).
This Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone ID8 is validated for use in IP, WB, IC, IH(P) for the detection of Myosin.

Physical form

Format: Purified

Analysis Note

Control
POSITIVE CONTROL: Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).

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

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Quality

Enter Lot Number to search for Certificate of Quality (COQ).

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