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MAB3568 Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone ID8

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MAB3568
100 µg  
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      Übersicht

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      B, H, Po, RbIP, WB, ICC, IH(P)MPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMAB3568
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone ID8
      Product Information
      FormatPurified
      Control
      • POSITIVE CONTROL: Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).
      PresentationLiquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationThis 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.
      Key Applications
      • Immunoprecipitation
      • Western Blotting
      • Immunocytochemistry
      • Immunohistochemistry (Paraffin)
      Application NotesWestern 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).
      Biological Information
      ImmunogenCrude smooth muscle extract from normal human adult uterus.
      Epitopesmooth muscle heavy chain, SM1 & SM2
      CloneID8 (also referred to as SMMS-1)
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificitySmooth muscle myosin, heavy chain. By Western blot the antibody recognizes a protein of 200-204 kDa.
      IsotypeIgG1
      Species Reactivity
      • Bovine
      • Human
      • Pig
      • Rabbit
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • CCDC128
      • MGC111781
      • FLJ16566
      UniProt Number
      UniProt SummarySIZE: 780 amino acids; 88314 Da
      Product Usage Statements
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsMaintain at 2-8*C in undiluted aliquots up to 6 months after date of receipt
      Packaging Information
      Material Size100 µg
      Global Trade Item Number
      Bestellnummer GTIN
      MAB3568 04053252505898