MRCL3 [p Ser19, p Ser20] Antibody
Novus Biologicals, part of Bio-Techne | Catalog # 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.
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] - 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] - 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] - 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
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
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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