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Primary Antibodies  >  Enzyme Antibodies
 

Myosin phospho S19/phospho S20 Antibody

Rabbit Polyclonal
Code

600-401-416
Size

100 µg
Price

$355.00
Availability

Ships next business day.
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Rockland's affinity purified anti-Monophosphorylated RLC Smooth and Non-Muscle Myosin pS19/20 antibody was used at 2.5 µg/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows strong staining of both vascular and myometrial smooth muscle cells of the uterus.  Tissue was formalin-fixed and paraffin embedded.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.  Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.
Affinity Purified Phospho specific antibody to Monophosphorylated Regulatory Light Chain of Smooth and Non-muscle Myosin at pS19/pS20 was used at a 1:5000 dilution to detect myosin light chain by Western blot. Either 13 or 20 ?l of a mouse cardiac myocyte lysate was loaded on a 4-20% Criterion gel for SDS-PAGE.  Samples were either mock-treated or CLA-treated, as indicated.  After washing, a 1:5,000 dilution of HRP conjugated Gt-a-Rabbit IgG (611-103-122) preceded color development using Amersham's substrate system. Other detection methods will yield similar results. Data courtesy of the Alliance for Cellular Signaling (http://www.signaling-gateway.org).
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Synonyms: Myosin regulatory light chain 12A Alternative name(s): Myosin regulatory light chain MRLC3 Myosin regulatory light chain 2, nonsarcomeric Myosin RLC MLC-2B

Myosin phospho S19/phospho S20 Antibody Properties

Description
Anti-Myosin pS19/pS20 (RABBIT) Antibody - 600-401-416
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human
Clonality
Polyclonal
Application
ELISA : 1:5,000 - 1:20,000
Western Blot : 1:1,000 - 1:5,000
Immunochemistry: 2.5 µg/ml
ImmunoPrecipitation: 1:100
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
MYL12A
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Lot#  

Myosin phospho S19/phospho S20 Antibody Description

Background
Myosin is the major component of thick muscle filaments, and is a long asymmetric molecule containing a globular head and a long tail.  The molecule consists of two heavy chains each ~200,000 daltons, and four light chains each ~16,000 - 21,000 daltons.  Activation of smooth and cardiac muscle primarily involves pathways that increase calcium levels and myosin phosphorylation, resulting in contraction.   Myosin light chain phosphatase acts to regulate muscle contraction by dephosphorylating activated myosin light chain.  This antibody is specific for the phosphorylated form of myosin light chain.  The selected peptide sequence used to generate the polyclonal antibody is located near the amino terminal end of the polypeptide corresponding to the smooth/non-muscle form of myosin regulatory light chain found in cardiac myocytes in addition to smooth and non-muscle cells.  This sequence differs from that of the sarcomeric/cardiac form of myosin regulatory light chain that has a different sequence around the phosphorylation site.  Human and mouse have almost identical sequences.  In human the phosphorylation site is pS19, while in mouse the site maps to pS20.
Immunogen
Human Myosin Light Chain phospho peptide corresponding to a region near the amino terminus of the human smooth/non-muscle form of myosin regulatory light chain conjugated to Keyhole Limpet Hemocyanin (KLH).
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This phospho specific polyclonal antibody was tested by ELISA, immunohistochemistry and immunoblotting.   Immunoblotting was used to show reactivity with unstimulated and stimulated cardiac myocytes.  The antibody was also reactive with the phosphorylated form of the immunizing peptide and minimally reactive with the non-phosphorylated form of the immunizing peptide.  Although not tested, this antibody is likely functional by immunoprecipitation.
Purity/Specifity
This affinity purified antibody is directed against the regulatory light chain of smooth and non-muscle myosin. The antibody is phosphospecific and detects monophosphorylated and diphosphorylated forms of the protein. The product was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide.  This phosphospecific polyclonal antibody is specific for the phosphorylated pS19/pS20 form of the protein, depending on the source origin of the protein.  Reactivity with non-phosphorylated myosin light chain is less than 1% by ELISA.  Cross reactivity is expected with myosin light chain from human and mouse.  Reactivity with the protein from other species has not been determined.  However, the sequence of the immunogen is nearly identical in mammalian and avian species.  BLAST search analysis was used to determine that the smooth and non-muscle forms of myosin regulatory light chain have identical sequences.  Cross reactivity is expected.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
DiGiovanna, M.P., Stern, D.F and Roussel (1996) Production of Pab anti-Phosphopeptide Antibodies. Curr. Prot. Immunol. 11; 6.1-6.19.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
human, mouse, rat
Catalog Number
600-401-905

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
human
Catalog Number
600-401-928

Product Type
Primary Antibodies;
Applications
ELISA, WB, IP
Reacts With
human, mouse, rat
Catalog Number
600-401-938

Product Type
Primary Antibodies;
Applications
ELISA, WB, IP
Reacts With
human, mouse, chimpanzee, rat
Catalog Number
600-401-959

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry