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MRCL3 [p Ser19, p Ser20] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NB100-1687

Novus Biologicals, part of Bio-Techne
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NB100-1687

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

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). (Uniprot: P19105)

Reactivity Notes

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.

Modification

p Ser19, p Ser20

Localization

Cytoplasmic

Specificity

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 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.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This affinity purified antibody is directed against the regulatory light chain of smooth and non-muscle myosin. 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

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for MRCL3 [p Ser19, p Ser20] Antibody

Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - This antibody has been shown to cross-react with mouse skeletal muscle regulatory light chain (Lai et al., 2016. Am J Physiol Endocrinol Metab), and is shown to positively react with mouse EDL muscle here. This image was submitted via customer Review.
Immunohistochemistry: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Immunohistochemistry: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Immunohistochemistry: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Used at 2.5 ug/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.
Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687]

Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Affinity purified phosphospecific antibody to monophosphorylated regulatory light chain of smooth and non-muscle Myosin at pS19 was used at a 1:1000 dilution to detect myosin light chain by Western blot. Approximately 12 ul of a mouse cardiac myocyte lysate was loaded on a 4-20% Criterion gel for SDS-PAGE. Samples were either mock treated or insulin or CLA treated according as indicated. After washing, a 1:5,000 dilution of HRP conjugated Gta- Rabbit IgG (611-103-122) preceded color development using Amersham's substrate system. Other detection methods will yield similar results.

Applications for MRCL3 [p Ser19, p Ser20] Antibody

Application
Recommended Usage

ELISA

1:10000-1:30000

Immunohistochemistry

2.5 ug/ml

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:100

Western Blot

1:500-1:2000
Application Notes
This product was tested by ELISA, immunohistochemistry, and western blotting. Immunoblotting was used to show reactivity with unstimulated and stimulated cardiac myocytes, 3T3 whole cell lysates, and regulatory light chain and smooth muscle phospho recombinant protein. 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.

Reviewed Applications

Read 1 review rated 5 using NB100-1687 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: MRCL3

The muscle myosins and hexomeric proteins consisting of two heavy chains and two pairs of light chains, the latter called essential (ELC) and regulatory (RLC). The light chains stabilize the long alpha helical neck of the myosin head (1). Human myosin light chain-2 (MYL2) is an important protein involved in the regulation of myosin ATPase activity in smooth muscle. In cardiac muscle, the precise role of MYL2 is not well understood; however, an increase in ventricular MYL2 is observed during myocardial hypertrophy in cardiac patients with valve stenosis (2). Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disease caused by mutations in all of the major sarcomeric proteins, including the ventricular myosin regulatory light-chain (RLC). The Myosin RLC mutation has been associated with a rare variant of cardiac hypertrophy defined by mid-left ventricular obstruction due to papillary muscle hypertrophy. This mutation was later found to cause ventricular and septal hypertrophy (3).

Alternate Names

MLCBMyosin RLC, MRCL3, MRLC3MLC-2B, MYL2B, myosin regulatory light chain 12A, Myosin regulatory light chain 2, nonsarcomeric, myosin regulatory light chain 3, myosin regulatory light chain MRCL3, Myosin regulatory light chain MRLC3, myosin, light chain 12A, regulatory, non-sarcomeric, myosin, light polypeptide, regulatory, non-sarcomeric (20kD), RLC

Entrez Gene IDs

4632 (Human)

Gene Symbol

MYL12A

UniProt

Additional MRCL3 Products

Product Documents for MRCL3 [p Ser19, p Ser20] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for MRCL3 [p Ser19, p Ser20] Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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