MAB3570
Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone N1/5
clone N1/5 (SM-M5), Chemicon®, from mouse
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About This Item
Codice UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Prodotti consigliati
Origine biologica
mouse
Livello qualitativo
Forma dell’anticorpo
purified immunoglobulin
Tipo di anticorpo
primary antibodies
Clone
N1/5 (SM-M5), monoclonal
Reattività contro le specie
rabbit, bovine, pig, human
Produttore/marchio commerciale
Chemicon®
tecniche
immunocytochemistry: suitable
immunohistochemistry: suitable (paraffin)
immunoprecipitation (IP): suitable
western blot: suitable
Isotipo
IgG1
N° accesso NCBI
N° accesso UniProt
Condizioni di spedizione
wet ice
modifica post-traduzionali bersaglio
unmodified
Informazioni sul gene
human ... MYH11(4629)
Applicazioni
Detect Myosin using this Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone N1/5 validated for use in IP, WB, IC, IH(P).
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. The antibody has also been used successfully on methyl-Carnoy fixed tissue.
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 MAB3570
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).
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. The antibody has also been used successfully on methyl-Carnoy fixed tissue.
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 MAB3570
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).
Stato fisico
Format: Purified
Risultati analitici
Control
POSITIVE CONTROL:
Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).
POSITIVE CONTROL:
Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).
Altre note
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Note legali
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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Codice della classe di stoccaggio
10 - Combustible liquids
Classe di pericolosità dell'acqua (WGK)
WGK 2
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
Certificati d'analisi (COA)
Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.
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