Flow cytometric analysis of MLC2v and MLC2a expression in MLC2v-transfected 293F cells. Untransfected (dashed line histogram) and human MLC2v-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Purified Mouse Anti-Human MLC2v (Cat. No. 565496, left panel) and Purified Mouse Anti-Human MLC2a (Cat. No. 565496, middle panel) followed by PE Goat Anti-Mouse Ig (Cat. No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. Western blot analysis of MLC2v expression (right panel). Lysate from MLC2v-transfected 293F cells was prepared for electrophoresis (SDS-PAGE) in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes and then probed with 0.063 (lane 1), 0.032 (lane 2), and 0.016 (lane 3) µg/mL of Purified Mouse Anti-MLC2v. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat No 554002).
Immunohistochemical staining of MLC2v in human left ventricle and left atrium. Following antigen retrieval with BD Retrievagen A buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 550878 ; top left) or Purified Mouse Anti-MLC2v (top right) on left ventricle. Additionally, Purified Mouse Anti-MLC2v (bottom left) and Purified Mouse Anti-MLC2a (Cat. No. 565496, bottom right) were used to stain left atrium. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Ig (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissues were counterstained with Hematoxylin. Original magnification: 20X (MLC2v) and 40X (MLC2a).
Flow cytometric analysis of MLC2v and MLC2a expression in MLC2v-transfected 293F cells. Untransfected (dashed line histogram) and human MLC2v-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Purified Mouse Anti-Human MLC2v (Cat. No. 565496, left panel) and Purified Mouse Anti-Human MLC2a (Cat. No. 565496, middle panel) followed by PE Goat Anti-Mouse Ig (Cat. No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. Western blot analysis of MLC2v expression (right panel). Lysate from MLC2v-transfected 293F cells was prepared for electrophoresis (SDS-PAGE) in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes and then probed with 0.063 (lane 1), 0.032 (lane 2), and 0.016 (lane 3) µg/mL of Purified Mouse Anti-MLC2v. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat No 554002).
Immunohistochemical staining of MLC2v in human left ventricle and left atrium. Following antigen retrieval with BD Retrievagen A buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 550878 ; top left) or Purified Mouse Anti-MLC2v (top right) on left ventricle. Additionally, Purified Mouse Anti-MLC2v (bottom left) and Purified Mouse Anti-MLC2a (Cat. No. 565496, bottom right) were used to stain left atrium. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Ig (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissues were counterstained with Hematoxylin. Original magnification: 20X (MLC2v) and 40X (MLC2a).
Product Details
BD Pharmingen™
MLC-2; MLC-2v; MYL2; MLC2; CMH10
Human (QC Testing), Mouse (Predicted)
Mouse IgG1, κ
Human MLC2v
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required), Western blot (Tested During Development)
28 kDa
0.5 mg/ml
AB_2739266
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Retrievagen A (pH 6.0) RUO
Cat No.
550524
Purified Mouse IgG1 κ Isotype Control RUO
Size
1 mL
Cat No.
550878
565497 Rev. 1
Antibody Details
S53-5
Myosin is composed of 2 heavy chains, 2 regulatory light chains, and 2 alkali light chains. The S53-5 monoclonal antibody specifically binds to human Myosin regulatory light chain 2,ventrical isoform (MLC2v) that is selectively expressed in the cardiac ventrical, while the MLC2a isoform is selectively expressed in the cardiac atrium. The differential expression of the two MLC2 isoforms is first detected early in cardiogenesis, prior to the formation of distinct atrial and ventrical chambers. During the induction of cardiovascular lineage cells from a human embryonic stem cell line, the appearance of contracting cardiomyocytes is coincident with the up-regulation of the expression of several genes, including the genes encoding MLC2a and MLC2v. Cross-reactivity of the S53-5 monoclonal antibody to mouse MLC2v is predicted due to the ~96% conservation of the human and mouse protein sequences.
565497 Rev. 1
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
565497 Rev.1
Citations & References
Development References (5)
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Dubois NC, Craft AM, Sharma P, et al. SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells. Nat Biotechnol. 2011; 29:1011-1018. (Biology). View Reference
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Franco D, Lamers WH, Moorman AF. Patterns of expression in the developing myocardium: towards a morphologically integrated transcriptional model. Cardiovasc Res. 1998; 38(1):25-53. (Biology). View Reference
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Laflamme MA, Murry CE. Heart regeneration. Nature. 2011; 473(7347):326-335. (Biology). View Reference
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Lee MY, Sun B, Schliffke S, et al. Derivation of functional ventricular cardiomyocytes using endogenous promoter sequence from murine embryonic stem cells. Stem Cell Res. 2012; 8(1):49-57. (Biology). View Reference
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O'Brien TX, Lee KJ, Chien KR. Positional specification of ventricular myosin light chain 2 expression in the primitive murine heart tube. Proc Natl Acad Sci U S A. 1993; 90(11):5157-5161. (Biology). View Reference
565497 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
