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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| B, H, Po, Rb | IP, WB, ICC, IH(P) | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB3570 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Myosin Antibody, smooth muscle heavy chain, SM1 & SM2, clone N1/5 |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Liquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | 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). |
| Key Applications |
|
| Application Notes | 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 its 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 | |
|---|---|
| Immunogen | Crude smooth muscle extract from normal human adult uterus. |
| Epitope | smooth muscle heavy chain, SM1 & SM2 |
| Clone | N1/5 (also referred to as SM-M5) |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Smooth muscle myosin, heavy chain. By Western blot the antibody recognizes a protein of 200-204 kDa. |
| Isotype | IgG1 |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | SIZE: 780 amino acids; 88314 Da |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at 2-8*C in undiluted aliquots up to 6 months after date of receipt |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |