LEFT PANEL: Western blot analysis of Cardiac Troponin T expression in human and mouse tissues. Human heart lysate was obtained from Abcam (Cat .No. ab2943, lane 1). C2C12 mouse myoblasts (ATCC CRL-1772) were cultured in DMEM containing 10% FBS (lane 2) or 0.5% FBS for 14 days for induction of cell differentiation (lane 3). Lysates were prepared, electrophoresed (SDS-PAGE) and transferred to PVDF membranes. The membranes were labeled with 1 µg/mL of Purified Mouse Anti-Cardiac Troponin T. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat. No. 554002). RIGHT PANEL: Immunohistochemical staining of Cardiac Troponin T in human heart. 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, left) or Purified Mouse Anti-Cardiac Troponin T (right). A three-step staining procedure that employs Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. Original magnification: 40×.
Flow cytometric analysis of Cardiac Troponin T in human (left panel) and mouse (right panel) cardiomyocytes differentiated in vitro. Human embryonic stem cell-derived cardiomyocytes (Evans lab, UCSD) were disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash buffer I (Cat. No. 557885). The C2C12 mouse myoblast cell line (ATCC CRL-1772) was cultured for 5 days in low-serum conditions for induction of cell differentiation, fixed with BD Cytofix™ Fixation Buffer, and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121, dashed line) or Purified Mouse Anti-Cardiac Troponin T (solid line). The second-step reagents were FITC Goat Anti-Mouse Ig (Cat. No. 554001, left) and PE Goat Anti-Mouse Ig (Cat. No. 550589, right). Events with the forward and side light-scatter characteristics of intact cells were gated. Flow cytometry was performed on a BD LSR Fortessa™ (left) or a BD FACSCanto™ II (right) flow cytometry system.
LEFT PANEL: Western blot analysis of Cardiac Troponin T expression in human and mouse tissues. Human heart lysate was obtained from Abcam (Cat .No. ab2943, lane 1). C2C12 mouse myoblasts (ATCC CRL-1772) were cultured in DMEM containing 10% FBS (lane 2) or 0.5% FBS for 14 days for induction of cell differentiation (lane 3). Lysates were prepared, electrophoresed (SDS-PAGE) and transferred to PVDF membranes. The membranes were labeled with 1 µg/mL of Purified Mouse Anti-Cardiac Troponin T. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat. No. 554002). RIGHT PANEL: Immunohistochemical staining of Cardiac Troponin T in human heart. 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, left) or Purified Mouse Anti-Cardiac Troponin T (right). A three-step staining procedure that employs Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. Original magnification: 40×.
Flow cytometric analysis of Cardiac Troponin T in human (left panel) and mouse (right panel) cardiomyocytes differentiated in vitro. Human embryonic stem cell-derived cardiomyocytes (Evans lab, UCSD) were disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash buffer I (Cat. No. 557885). The C2C12 mouse myoblast cell line (ATCC CRL-1772) was cultured for 5 days in low-serum conditions for induction of cell differentiation, fixed with BD Cytofix™ Fixation Buffer, and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121, dashed line) or Purified Mouse Anti-Cardiac Troponin T (solid line). The second-step reagents were FITC Goat Anti-Mouse Ig (Cat. No. 554001, left) and PE Goat Anti-Mouse Ig (Cat. No. 550589, right). Events with the forward and side light-scatter characteristics of intact cells were gated. Flow cytometry was performed on a BD LSR Fortessa™ (left) or a BD FACSCanto™ II (right) flow cytometry system.
Product Details
BD Pharmingen™
cTnT, Cardiac Muscle Troponin T, Troponin T type 2, TNNT2
Mouse (QC Testing), Human (Tested in Development), Rat,Pig,Dog,Chicken,Rabbit,Guinea Pig (Reported)
Mouse BALB/c IgG1, κ
Rabbit cardiac troponin T Protein
Intracellular staining (flow cytometry) (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required), Western blot (Tested During Development)
35-40 kDa
0.5 mg/ml
AB_2738938
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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
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/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Retrievagen A (pH 6.0) RUO
Cat No.
550524
564766 Rev. 1
Antibody Details
13-11
The 13-11 monoclonal antibody specifically recognizes Troponin T type 2 (cardiac), encoded by the gene TNNT2. Troponin T is the tropomyosin-binding subunit of the troponin complex, which also encompasses troponin C and troponin I. This complex regulates muscle contraction in skeletal and cardiac muscle in response to alterations in calcium levels. Troponin T type 2 is solely found in the heart, and genetic alterations in the TNNT2 gene are associated to a series of heart disorders in humans, including hypertrophic cardiomyopathy, dilated cardiomyopathy and left ventricular noncompaction. Cardiac Troponin T can be used as a marker for the identification of cardiomyocytes derived from pluripotent stem cells.
564766 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
564766 Rev.1
Citations & References
View product citations for antibody "564766" on CiteAb
Development References (7)
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Jáchymová M, Muravská A, Paleček T, et al. Genetic variation screening of TNNT2 gene in a cohort of patients with hypertrophic and dilated cardiomyopathy. Physiol Rev. 2012; 61(2):169-175. (Biology). View Reference
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Lian X, Hsiao C, Wilson G, et al. Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A. 2012; 109(27):E1848-E1857. (Clone-specific: Flow cytometry). View Reference
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Malouf NN, McMahon D, Oakeley AE, Anderson PA. A cardiac troponin T epitope conserved across phyla. J Biol Chem. 1992; 267(13):9269-9274. (Immunogen: Electron microscopy, ELISA, Immunofluorescence, Western blot). View Reference
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Mauritz C, Schwanke K, Reppel M, et al. Generation of functional murine cardiac myocytes from induced pluripotent stem cells. Circulation. 2008; 118(5):507-517. (Clone-specific: Immunofluorescence). View Reference
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Thierfelder L, Watkins H, MacRae C, et al. Alpha-tropomyosin and cardiac troponin T mutations cause familial hypertrophic cardiomyopathy: a disease of the sarcomere.. Cell. 1994; 77(5):701-12. (Biology). View Reference
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Uosaki H, Fukushima H, Takeuchi A, et al. Efficient and scalable purification of cardiomyocytes from human embryonic and induced pluripotent stem cells by VCAM1 surface expression. PLoS ONE. 6(8)(Clone-specific: Flow cytometry). View Reference
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Zhang J, Wilson GF, Soerens AG, et al. Functional cardiomyocytes derived from human induced pluripotent stem cells. Circ Res. 104(4)(Clone-specific: Immunofluorescence). View Reference
564766 Rev. 1
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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.