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48243
Cardiogenesis Marker Antibody Sampler Kit

Cardiogenesis Marker Antibody Sampler Kit #48243

IF-IC Image 1

Confocal immunofluorescent analysis of HEK293 (positive, left) and PANC-1 (negative, right) cells, using NKX2.5 (E1Y8H) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 2

Western blot analysis of extracts from various cell lines using NKX2.5 (E1Y8H) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Chromatin IP-seq Image 3

Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across FGFR2, a known target gene of GATA-6 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 4

Western blot analysis of extracts from Huh7 and 293 cells using GATA-6 (D61E4) XP® Rabbit mAb.

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Western Blotting Image 5

Western blot analysis of extracts from various cells lines using MEF2C (D80C1) XP® Rabbit mAb.

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Western Blotting Image 6

Western blot analysis of extracts from various cell lines using α-Actinin (D6F6) XP® Rabbit mAb.

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Western Blotting Image 7

Western blot analysis of extracts from human, mouse and rat heart tissue using Troponin I (D6F8) Rabbit mAb.

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Western Blotting Image 8

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a human cardiac Troponin T (isoform 1) construct (+), using Troponin T (Cardiac) Antibody.

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Western Blotting Image 9

Western blot analysis of extracts from untreated and PMA-treated C6, COS, HeLa and C2C12 cells using Connexin 43 Antibody.

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Western Blotting Image 10

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Flow Cytometry Image 11

Flow cytometric analysis of PANC-1 cells (blue) and HEK293 cells (green) using NKX2.5 (E1Y8H) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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Flow Cytometry Image 12

Flow cytometric analysis of SKOV3 cells (blue) and HUH-7 cells (green) using GATA-6 (D61E4) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.

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Chromatin IP Image 13

Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and either GATA-6 (D61E4) XP® Rabbit mAb, or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human FDPS Promoter Primers #13840, human FGFR2 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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IF-IC Image 14

Confocal immunofluorescent analysis of C2C12 cells, undifferentiated (left) or differentiated for 3 days (right), using MEF2C (D80C1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

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IF-IC Image 15

Confocal immunofluorescent analysis of SNB19 cells using α-Actinin (D6F6) XP® Rabbit mAb (green) showing colocalization with actin filaments that were labeled with β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 16

Western blot analysis of extracts from human and rat heart and human skeletal muscle using Troponin T (Cardiac) Antibody.

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Connexin 43 Antibody.

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IF-IC Image 18

Confocal immunofluorescent analysis of KM12 (left) and SK-OV-3 cells (right) using GATA-6 (D61E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 19

Immunohistochemical analysis of paraffin-embedded human heart, using Connexin 43 Antibody. (High magnification, inset).

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Connexin 43 Antibody.

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IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded mouse heart using Connexin 43 Antibody in the presence of control peptide (left) or antigen specific peptide (lright).

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IHC-F (frozen) Image 22

Immunohistochemical analysis of frozen mouse heart tissue, using Connexin 43 Antibody.

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IF-IC Image 23

Confocal immunofluorescent analysis of confluent COS cells using Connexin 43 Antibody (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-F Image 24

Confocal immunofluorescent analysis of rat optic nerve and ciliary epithelium using Connexin 43 Antibody (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

See notes for additional testing details.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
NKX2.5 (E1Y8H) Rabbit mAb 8792 20 µl
  • WB
  • IF
  • F
H 30-42 Rabbit IgG
GATA-6 (D61E4) XP® Rabbit mAb 5851 20 µl
  • WB
  • IF
  • F
  • ChIP
H 55 Rabbit IgG
MEF2C (D80C1) XP® Rabbit mAb 5030 20 µl
  • WB
  • IP
  • IF
H M 50-60 Rabbit IgG
α-Actinin (D6F6) XP® Rabbit mAb 6487 20 µl
  • WB
  • IF
H M R Mk 100 Rabbit IgG
Troponin I (D6F8) Rabbit mAb 13083 20 µl
  • WB
H M R 28 Rabbit IgG
Troponin T (Cardiac) Antibody 5593 20 µl
  • WB
H R 40 Rabbit 
Connexin 43 Antibody 3512 20 µl
  • WB
  • IHC
  • IF
H M R Mk Z 39, 41, 43, 44 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Cardiogenesis Marker Antibody Sampler Kit provides an economical means of evaluating proteins involved in heart development. This kit contains enough antibody to perform two western blot experiments per primary antibody.

NKX2.5 (E1Y8H) Rabbit mAb recognizes endogenous levels of total NKX2.5 protein. GATA-6 (D61E4) XP® Rabbit mAb recognizes endogenous levels of total GATA-6 protein. MEF2C (D80C1) XP® Rabbit mAb detects endogenous levels of total MEF2C protein. α-Actinin (D6F6) XP® Rabbit mAb recognizes endogenous levels of total α-actinin protein. Troponin I (D6F8) Rabbit mAb recognizes endogenous levels of total troponin I protein. Troponin T (Cardiac) Antibody detects endogenous levels of total cardiac Troponin T protein. Connexin 43 Antibody detects endogenous levels of total connexin 43. This antibody does not cross-react with other connexins.

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro67 of human NKX2.5 protein, residues near the amino terminus of human GATA-6 protein, a region surrounding Met182 of human MEF2C protein, residues surrounding Phe316 of human α-actinin protein, or residues near the amino terminus of human troponin I protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to a region surrounding Pro69 of human cardiac troponin T protein or residues of human connexin 43. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Cardiogenesis is a complex developmental event involving numerous transcription factors. NKX2.5 is a member of the NKX homeobox transcription factor family, which plays an essential role in heart development and is among the earliest factors expressed in the cardiac lineage in developing embryos. Mutations in NKX2.5 are associated with several congenital heart conditions, such as atrial defect with atrioventricular conduction defects (ASD-AVCD) and Tetralogy of Fallot (TOF) (1,2). GATA proteins comprise a group of transcription factors that are related by the presence of conserved zinc finger DNA binding domains, which bind directly to the nucleotide sequence core element GATA (3-5). GATA-6 plays a critical role in endoderm development and knock out of GATA-6 is embryonic lethal due to defects in formation of the heart tube and a failure to develop extraembryonic endoderm (6). MEF2C is a member of the MEF2 (myocyte enhancer factor 2) family of transcription factors. The MEF2 family members were originally described as muscle-specific DNA binding proteins that recognize MEF2 motifs found within the promoters of many muscle-specific genes (7,8). α-Actinin was first recognized as an actin cross-linking protein. The α-actinin protein interacts with a large number of proteins involved in signaling to the cytoskeleton, including those involved in cellular adhesion, migration, and immune cell targeting (9). The muscle isoforms 2 and 3 (ACTN2, ACTN3) localize to the Z-discs of striated muscle and to dense bodies and plaques in smooth muscle (9). Troponin, working in conjunction with tropomyosin, functions as a molecular switch that regulates muscle contraction in response to changes in the intracellular Ca2+ concentration. Troponin consists of three subunits: the Ca2+-binding subunit troponin C (TnC), the tropomyosin-binding subunit troponin T (TnT), and the inhibitory subunit troponin I (TnI) (10). Assays for measuring serum concentrations of cardiac muscle TnT (cTNT), as well as cTnI, have been reported for analyzing cardiac injury. Connexin 43 (Cx43) is a member of the large family of gap junction proteins, which assemble as a hexamer and are transported to the plasma membrane to create a hemichannel that can associate with hemichannels on nearby cells to create cell-to-cell channels. Gap junction communication is important in development and regulation of cell growth. Phosphorylation of Cx43 is important in regulating assembly and function of gap junctions (11,12).

  1. Ko, L.J. and Engel, J.D. (1993) Mol Cell Biol 13, 4011-22.
  2. Otey, C.A. and Carpen, O. (2004) Cell Motil Cytoskeleton 58, 104-11.
  3. Ward, D.G. et al. (2002) J. Biol. Chem. 277, 41795-41801.
  4. Musil, L.S. et al. (1990) J Cell Biol 111, 2077-88.
  5. Merika, M. and Orkin, S.H. (1993) Mol Cell Biol 13, 3999-4010.
  6. Martin, J. F. et al. (1994) Mol. Cell. Biol. 14, 1647-1656.
  7. Musil, L.S. and Goodenough, D.A. (1991) J Cell Biol 115, 1357-74.
  8. Benson, D.W. et al. (1999) J Clin Invest 104, 1567-73.
  9. Lowry, J.A. and Atchley, W.R. (2000) J Mol Evol 50, 103-15.
  10. Yu, Y. T. et al. (1992) Genes Dev. 6, 1783-1798.
  11. Reamon-Buettner, S.M. and Borlak, J. (2010) Hum Mutat 31, 1185-94.
  12. Cai, K.Q. et al. (2008) Dev Dyn 237, 2820-9.
Entrez-Gene Id
87 , 2697 , 2627 , 4208 , 1482 , 7137
Swiss-Prot Acc.
P12814 , P17302 , Q92908 , Q06413 , P52952 , P19429
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.

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