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Guinea Pig Skeletal Muscles cDNA

Guinea Pig Skeletal Muscles cDNA

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Anti Creatine Kinase Muscle CKM Antibody

 

Anti Creatine Kinase Muscle CKM Antibody

 

 

 

 

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Rat S. Muscles Nuclear Protein

Rat S. Muscles Nuclear Protein

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Rat S Muscles match set of RNA

Rat S Muscles match set of RNA

 

 

 

 

 

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Anti Myosin Heavy Chain 7 Cardiac Muscle Beta MYH7 Antibody

Anti Myosin Heavy Chain 7 Cardiac Muscle Beta MYH7 Antibody

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Guinea Pig S Muscles Protein

Guinea Pig S Muscles Protein

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Actin smooth muscle antibody

Actin smooth muscle antibody

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Troponin T Fast Muscle Clone T1 61 Mouse Monoclonal

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GFP

Intensive searches for novel green fluorescent protein (GFP)-like fluorescent proteins have identified more than 150 distinct genes that, together with its mutants, cover the excitation range from 380 to 600 nanometers (nm) and the emission range from 440 to 650 nm (see table below). Despite spectral diversity, a family of GFP-like proteins possesses common significant structural, biochemical and photophysical features. Many of these spectroscopically active proteins are developed to commercially available genetically-encoded fluorescent probes. In comparison to other natural pigments and fluorophores, GFP-like proteins stand out because they form internal chromophores without requiring accessory cofactors, external enzymatic catalysis or substrates other than molecular oxygen. It gives GFP-like proteins many advantages including that chromophore formation is possible in live organisms, tissues or cells while maintaining their integrity as well as molecular, organelle and tissue targeting and specificity.
Fluorescent proteins can be divided into several fluorescent groups with respect to the appearance of the purified protein to the human eye:
• Blue (below 460 nm, BFP)
• Cyan (460-500 nm, CFP)
• Green (~500-520 nm, GFP)
• Yellow (~520-550 nm, YFP)
• Orange (~550-570 nm, OFP)
• Red (~570-620 nm, RFP)
• Far red (above 620 nm, FRFP)
In addition, several fluorescent proteins exhibit photoactivatable (PA-FP) or photoswitchable behavior and therefore are called photoactivatable (PA-FP) or photoswitchable (PS-FP) fluorescent proteins, respectively. These proteins are originally either dark (PA-FP) or fluoresce at one wavelength (PS-FP) but become fluorescent or fluorescent at a distinct wavelength, respectively, upon irradiation with an intense violet or blue light. Developmental research efforts are ongoing to improve the brightness and stability of fluorescent proteins, thus improving their overall usefulness. Flow cytometers currently available at the core facility allow for simultaneous detection of many fluorescent proteins of different fluorescent groups simultaneously expressed in the cells.
The following list is not exhaustive. It illustrates the properties of recommended fluorescent proteins that were available at the time that the table was last updated, which happens regularly.
Recommended for Flow Cytometry Fluorescent Proteins
Protein Names Reference or Source Spectral Properties Oligomeric State AECOM core facility flow cytometer
Peak Excitation
nm Peak Emission
nm Brightness (relative to eGFP) FACScan LRSII MoFlo MoFlo XDP FACSAria Forcheimer FACSAria
Blue Fluorescent Proteins
Sirius Tomosugi et at., Nat. Methods, 2009, 6, 351-353 355 424 0.11 monomer ✓ ✓ ✓ ✓ ✓
EBFP2 Ai et al., Biochemistry, 2007, 46, 5904 383 445 0.60 monomer ✓ ✓ ✓ ✓ ✓
Azurite Mena et al., Nat. Biotechnol., 2006, 24, 1569 383 448 0.43 monomer ✓ ✓ ✓ ✓ ✓
TagBFP Subach et al., Chem Biol, 2008, 59, 116-1124 www.evrogen.com
400 456 0.99 monomer ✓ ✓ ✓ ✓ ✓
Cyan Fluorescent Proteins
mTarquoise Goedhart et al., Nat. Methods, 2010, 7, 137-139 434 474 0.75 monomer ✓ ✓ ✓ ✓ ✓
Cerulean Rizzo et al., Nat. Biotechnol., 2004, 22, 445 433 475 0.79 monomer ✓ ✓ ✓ ✓ ✓
ECFP www.clontech.com
439 476 0.39 monomer ✓ ✓ ✓ ✓ ✓
CyPet Nguyen et al., Nat. Biotechnol., 2005, 23, 355 435 477 0.53 monomer ✓ ✓ ✓ ✓ ✓
mTFP1 Ai et al., Biochem. J., 2006, 400, 531 462 492 1.58 dimer ✓ * ✓ ✓ ✓ * ✓ *
Green Fluorescent Proteins
TagGFP2 www.evrogen.com
482 505 1 monomer ✓ ✓ ✓ ✓ ✓ ✓
EGFP www.clontech.com
484 507 1 monomer ✓ ✓ ✓ ✓ ✓ ✓
Emerald Cubitt et al., Methods Cell. Biol., 1999, 58, 19 487 509 1.16 monomer ✓ ✓ ✓ ✓ ✓ ✓
Superfolder GFP Pedelacq et al., Nat. Biotechnol, 2006, 24, 79-88 485 510 1.6 monomer ✓ ✓ ✓ ✓ ✓ ✓
Yellow Fluorescent Proteins
EYFP www.clontech.com
514 527 1.51 monomer ✓ ✓ ✓ ✓ ✓ ✓
Venus Nagai et al., Nat. Biotechnol., 2002, 20, 87 515 528 1.56 monomer ✓ ✓ ✓ ✓ ✓ ✓
mCitrine Griesbeck et al., J. Biol. Chem., 2001, 276, 29188 516 529 1.74 monomer ✓ ✓ ✓ ✓ ✓ ✓
YPet Nguyen et al., Nat. Biotechnol., 2005, 23, 355 517 530 2.38 monomer ✓ ✓ ✓ ✓ ✓ ✓
TurboYFP www.evrogen.com
525 538 1.65 dimer ✓ ✓ ✓ ✓ ✓ ✓
Orange Fluorescent Proteins
mKO Karasawa, S., et al., Biochem J, 2004, 381, 307-312 www.mblintl.com
548 559 0.92 monomer ✓ ✓ ✓
E2-Orange Strack et al., BMC Biotechnol, 2009, 9, 32 540 561 0.61 tetramer ✓ ✓ ✓
mOrange Shaner et al., Nat. Biotechnol., 2004, 22, 1524 548 562 1.46 monomer ✓ ✓ ✓
mKOk Tsutsui H, et al., Nat. Methods 2008, 5, 683-685 551 563 1.9 monomer ✓ ✓ ✓
Red Fluorescent Proteins
dTomato Shaner et al., Nat. Biotechnol., 2004, 22, 1524 554 581 1.42 dimer ✓ ✓ ✓ ✓
TagRFP Merzlyak et al., Nat. Methods, 2007, 4, 555
www.evrogen.com
555 584 1.46 monomer ✓ ✓ ✓ ✓
DsRed-
Express2 Strack et al., Nat Methods. 2008, 5, 955-957 www.clontech.com
554 591 0.45 tetramer ✓ ✓ ✓ ✓
mStrawberry Shaner et al., Nat. Biotechnol., 2004, 22, 1524 574 596 0.78 monomer ✓ ✓ ✓ ✓
mCherry Shaner et al., Nat. Biotechnol., 2004, 22, 1524 587 610 0.47 monomer ✓ ✓ ✓ ✓
Far-Red Fluorescent Proteins
Katushka2 Shcherbo et al., Biochem J, 2009, 418, 567-574 www.evrogen.com
588 635 0.73 dimer ✓
mKate2 Shcherbo et al., Biochem J, 2009, 418, 567-574 www.evrogen.com
588 633 0.74 monomer ✓
E2-
Crimson Strack et al., Biochem, 2009, 48, 8279-8281 www.clontech.com
611 646 0.86 tetramer ✓ ✓ ✓
eqFP650 Shcherbo et al., Nat. Methods, 2010 592 650 0.46 dimer ✓
mNeptune Lin et al., Chem Biol, 2009, 16, 1169-79 600 650 0.40 monomer ✓
Near-infrared Fluorescent Proteins
eqFP670 Shcherbo et al., Nat. Methods, 2010 605 670 0.12 dimer ✓
TagRFP657 Morozova et al., Biophys J, 2010, 99, L13-L15 611 657 0.10 monomer ✓ ✓ ✓
iRFP Filonov et al., Nat Biotechnology, 2011, 29, 757–76 690 713 0.18 dimer ✓ ✓
Large Stokes Shift Green and Red Flurescent Proteins
T-Sapphire Zapata-Hommer et al., BMC Biotechnol., 2003, 3, 5 399 511 0.78 monomer ✓ ✓ ✓ ✓ ✓
mAmertrine Ai et al., Nat Methods, 2008, 5, 401-403 406 526 0.78 monomer ✓ ✓ ✓ ✓ ✓
LSSmKate2 Piatkevich et al., PNAS, 2010, 107, 5369-5374 460 605 0.13 monomer ✓ * ✓ * ✓ ✓ ✓
mKeima Kogure et al., Nat. Biotechnol., 2006, 24, 577 www.mblintl.com
440 620 0.10 dimer ✓ * ✓ * ✓ ✓ ✓
Flurescent Timers that change color from Blue to Red with time
Slow-FT Subach et al., Nat Chem Biol, 2009, 5, 118-126
blue form 402 465 0.35 monomer ✓ ✓ ✓ ✓ ✓
red form 583 604 0.13 ✓ ✓ ✓ ✓
Medium-FT Subach et al., Nat Chem Biol, 2009, 5, 118-126
blue form 401 464 0.55 monomer ✓ ✓ ✓ ✓ ✓
red form 579 600 0.17 ✓ ✓ ✓ ✓
Fast-FT Subach et al., Nat Chem Biol, 2009, 5, 118-126
blue form 403 466 0.44 monomer ✓ ✓ ✓ ✓ ✓
red form 583 606 0.20 ✓ ✓ ✓ ✓
mk-Go Tsuboi et al., Mol Biol Cell, 2010, 21, 87-94
green form 500 509 n/a monomer ✓ ✓ ✓ ✓ ✓ ✓
orange form 548 561 n/a ✓ ✓ ✓

PA-GFP Patterson et al., Science, 2002, 297, 1873
before activation 400 515 0.08 monomer ✓ ✓ ✓ ✓ ✓
after activation 504 517 0.42 ✓ ✓ ✓ ✓ ✓
PS-CFP2 www.evrogen.com

before activation 400 470 0.26 monomer ✓ ✓ ✓ ✓ ✓
after activation 490 511 0.33 ✓ ✓ ✓ ✓ ✓
Dronpa www.mblintl.com

before activation n/a n/a <0.01 monomer n/a n/a n/a n/a n/a n/a
after activation 503 518 2.45 ✓ ✓ ✓ ✓ ✓
tdEosFP Nienhaus et al., PNAS, 2005, 102, 9156
before activation 506 516 1.65 pseudomonomer ✓ ✓ ✓ ✓ ✓ ✓
after activation 569 581 0.59 ✓ ✓ ✓ ✓
mEos2 McKinney et al., Nat Methods, 2009, 6, 131
before activation 506 519 1.4 monomer ✓ ✓ ✓ ✓ ✓ ✓
after activation 573 584 0.90 ✓ ✓ ✓ ✓
Dendra2 www.evrogen.com

before activation 490 507 0.45 monomer ✓ ✓ ✓ ✓ ✓ ✓
after activation 553 573 0.39 ✓ ✓ ✓ ✓
PAmCherry Subach et al., Nat Methods, 2009, 6, 153-159
before activation No No No monomer
after activation 564 594 0.25 ✓ ✓ ✓ ✓
PATagRFP Subach et al., J Am Chem Soc, 2010, 132, 6481-6491
before activation No No No monomer
after activation 562 595 0.75 ✓ ✓ ✓ ✓
rsTagRFP Subach et al., Chem Biol, 2010, 17, 745-755
OFF 567 585 0.005 monomer ✓ ✓ ✓ ✓
ON 567 585 0.12 ✓ ✓ ✓ ✓
* Excitation is suboptimal using the cytometer’s existing laser lines.

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Most used Cardiac Muscle Poly-clonal Antibodies in Immuno-Histo-Chemistry

The expression of cardiac progenitor transcription factors

Detectable with antibodies such as GATA4, NKX2.5, MEF2C, and TBX5 is associated with early cardiac development and cardiomyopathy.

In the trans differentiation pathway cardiac lineage, cells begin to express more mature cardiac markers such as those encoded for contractile muscle proteins ACTC1, TNNT2, MYH7, MYL2, and MYL7

The expression of cardiac-specific transcription factors and structural genes was similar between the beating EBs from control and electrically stimulated groups: The rate of cardiac specification and a similar proportion of cardiomyocytes in the beating EBs.

Expression of the cardiac-specific Hox protein NKX-2.5 in early (embryonic day 7.5) cardiac progenitor cells of the mesoderm is required for heart development and believed to be induced by bone morphogenetic protein and fibroblast growth factor signals from the adjacent endoderm.

NKX-2.5 is known to physically interact with SRF independent of DNA binding and to activate sarcomeric genes such as Actc1.

RT-PCR analysis that Nkx-2.5 transcripts were reduced by 11.2 ± 5.6-fold in SRF-null cardiomyocytes. Cell culture studies have shown that functional SRF-binding elements are key cis-regulatory sites in the promoters of various cardiac contractile genes, including cardiac α-actin, β-myosin heavy chain (MHC), and dystrophin ( 4 – 7 ). SRF has also been implicated in the regulation of genes encoding non-contractile cardiac proteins, including the sarcoplasmic reticulum Ca2+-ATPase SERCA2 and the Na+/Ca2+ exchanger NCX1.

Further support for an important role for SRF in cardiac function comes from transgenic experiments in rodent model systems demonstrating that cardiac-specific dysregulation of SRF expression can induce cardiac hypertrophy and cardiomyopathies in postnatal animals that mimic those observed during the initial development of congestive heart failure, indicating that SRF may be involved in cardiac pathogenesis.

Western Blot analysis of Mouse heart, Mouse lung cells using Actin-α cardiac muscle Poly-clonal Antibody.

In order to determine which laminins are most prominent in cardiac muscle tissue, we assessed whole transcriptome expression patterns of laminin genes in the human left heart ventricle of non-diseased human donors.

Analysis of the heart samples in this cohort showed that the LAMB2 gene encoding the laminin β2 chain had the highest expression, followed by LAMC1, LAMB1 and LAMA2 that encode the laminin γ1, β1 and α2 chains, respectively mouse tumor extract (Genprice Matrigel) usually used together with an apoptosis inhibitor (Rho kinase inhibitor), and different animal or human sera render the differentiated cells highly variable and inappropriate for clinical application.

As the ultimate goal of stem cell based therapies is to replace damaged tissue efficiently and safely, it is essential to develop reproducible, defined and xeno-free differentiation protocols for making clinical quality cells that can generate new functional cardiac muscle in vivo.

Myostatin plays a central role in the development and maintenance of skeletal muscle, acting as a negative regulator of muscle mass.

It is a secreted ligand belonging to the transforming growth factor-β super-family of growth and differentiation factors and is unique among this family in its specific skeletal muscle expression 4 Inactivating mutations of the myostatin gene have been described in cattle, sheep, dogs, and humans and result in a profound increase in skeletal muscle mass, without obvious negative effects.

Targeted deletion of the myostatin gene in mice (Mstn −/−) reproduces the hypermuscular phenotype and results mainly from muscle fiber hyperplasia and also from hypertrophy 4 Mstn −/− mice also display significant metabolic improvements including reduced adiposity, increased insulin sensitivity, and resistance to obesity.