![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.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Western-Blot-NB100-1687-img0019.jpg "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.")
| Reactivity | Hu, MuSpecies Glossary |
| Applications | WB, ELISA, IHC, IP |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| 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. |
| 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) |
| 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. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | MYL12A |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| 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. |
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| Reviewed Applications |
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| Storage | Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles. |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Preservative | 0.01% Sodium Azide |
| Purity | Immunogen affinity purified |
![MRCL3 [p Ser19, p Ser20] Antibody](/sites/all/modules/enterprise-tech/et_datasheets/images/novus_guarantee.png "MRCL3 [p Ser19, p Ser20] Antibody")
| Images | Ratings | Applications | Species | Date | Details | ||||||||
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reviewed by:
Verified Customer |
WB | Mouse | 05/30/2017 |
Summary
Comments
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Secondary Antibodies |
Isotype Controls |
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![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.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Immunohistochemistry-NB100-1687-img0015.jpg "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] - 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.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Western-Blot-NB100-1687-img0014.jpg "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.")
![Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - 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 preceded color development using Amersham's substrate system.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Western-Blot-NB100-1687-img0016.jpg "Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - 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 preceded color development using Amersham's substrate system.")
![Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Lane 1: Regulatory Light Chain Non-Phospho recombinant protein. Lane 2: Regulatory Light Chain Phospho recombinant protein. Lane 3: Smooth Muscle Non-Phospho recombinant protein. Lane 4: Smooth Muscle Phospho recombinant protein. Load: 50 ng per lane. Primary antibody: Myosin pS19/pS20 primary antibody at 1:1,000 overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 60 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 20 kDa, 20 kDa for Regulatory Light Chain Phospho. Other band(s): None.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Western-Blot-NB100-1687-img0017.jpg "Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Lane 1: Regulatory Light Chain Non-Phospho recombinant protein. Lane 2: Regulatory Light Chain Phospho recombinant protein. Lane 3: Smooth Muscle Non-Phospho recombinant protein. Lane 4: Smooth Muscle Phospho recombinant protein. Load: 50 ng per lane. Primary antibody: Myosin pS19/pS20 primary antibody at 1:1,000 overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 60 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 20 kDa, 20 kDa for Regulatory Light Chain Phospho. Other band(s): None.")
![Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Used at a 1:1000 dilution to detect myosin light chain by Western blot on 3T3 cell lysates. A standard urea/glycerol gel without SDS was used to separate phospho forms of regulatory light chain according to mass to charge ratios. In Panel A, reactivity of phosphospecific antibody is shown. In Panel B, reactivity of commercially available pan reactive antibody that detects both unphosphorylated and phosphorylated forms of regulatory light chain is shown. phosphospecific antibody detects both monophosphorylated (pSer20 Mono-P-RLC) and diphosphorylated (pThr19-pSer20 Di-P-RLC) regulatory light chain.](http://images.novusbio.com/fullsize/MRCL3-[p-Ser19--p-Ser20]-Antibody-Western-Blot-NB100-1687-img0018.jpg "Western Blot: MRCL3 [p Ser19, p Ser20] Antibody [NB100-1687] - Used at a 1:1000 dilution to detect myosin light chain by Western blot on 3T3 cell lysates. A standard urea/glycerol gel without SDS was used to separate phospho forms of regulatory light chain according to mass to charge ratios. In Panel A, reactivity of phosphospecific antibody is shown. In Panel B, reactivity of commercially available pan reactive antibody that detects both unphosphorylated and phosphorylated forms of regulatory light chain is shown. phosphospecific antibody detects both monophosphorylated (pSer20 Mono-P-RLC) and diphosphorylated (pThr19-pSer20 Di-P-RLC) regulatory light chain.")
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![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")
