human adult cfs  (PromoCell)

Journal: Molecular Biology International

Article Title: Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4 α Isoform in Humans

doi: 10.1155/2016/3105478

Figure Lengend Snippet: 2D gels of commercially available extracts of human adult heart. (a) The Coomassie stained human adult heart protein across the gel. Seven spots A, B, C, D, E, F, and G were identified, which are not visible clearly. (b) This panel represents the enlarged portion of the gel in (a) and shows clearly A–E spots. (c) The PVDF filter was stained with CH1 monoclonal antibody followed by treatment with secondary antibody as mentioned under Materials and Methods and subsequently treated with ECL and exposed to X-ray film. (d) Developed X-ray film was merged on the top of the Coomassie stained blot. (e) Enlarged nine spots (A–I) were marked on the PVDF filter (as in (a) and (b)) and on the duplicate gel. All the spots were marked and excised and were used for extraction of protein for subsequent mass spectrometric analyses. Extracted peptides from spots E, F, and G were analyzed for mass spectrometry as described in the method section.

Article Snippet: Human adult and fetal cardiac tissue RNA were obtained from Zyagen (San Diego, CA); adult human heart protein samples were obtained from Imgenex (San Diego, CA).

Techniques: Staining, Extraction, Mass Spectrometry

Journal: Molecular Biology International

Article Title: Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4 α Isoform in Humans

doi: 10.1155/2016/3105478

Figure Lengend Snippet: Expression of TPM4 isoforms with exon 9a/b in human heart and skeletal muscle by conventional RT-PCR. (a) Expression of TPM4α and TPMδ in human fetal and adult hearts as determined . TPM4 δ amplified with primer 1(+) and primer 2(−) as shown in and . The size of the amplified DNA is ~747 bp. Lane 1: adult heart; lane 2: fetal heart; lane 3: primer control. TPM4 α amplified with primer 1(+) and primer 2(−) as shown in and . The size of the amplified DNA is ~855 bp. Lane 4: adult heart; lane 5: fetal heart; lane 6: primer control . (b) Expression of TPM4α in human fetal heart , adult heart , and adult skeletal muscle . TPM4 α amplified with primer 1(+) and primer 2(−) as shown in and . The size of the amplified DNA is ~855 bp. Lane 1: fetal heart; lane 2: adult heart; lane 3: adult skeletal muscle (source 1); lane 4: adult skeletal muscle (source 2); lane 5: primer control.

Article Snippet: Human adult and fetal cardiac tissue RNA were obtained from Zyagen (San Diego, CA); adult human heart protein samples were obtained from Imgenex (San Diego, CA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Control

Journal: Molecular Biology International

Article Title: Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4 α Isoform in Humans

doi: 10.1155/2016/3105478

Figure Lengend Snippet: Relative expression of TPM4 α and TPM4 δ in human adult and fetal hearts. Fold changes of TPM4 α and TPM4 δ in human adult heart, fetal heart, and skeletal muscle. qRT-PCR was carried out in triplicate with isoform specific primer-pairs sequences of which are given in . (a) TPM4 α and (b) TPM4 δ .

Article Snippet: Human adult and fetal cardiac tissue RNA were obtained from Zyagen (San Diego, CA); adult human heart protein samples were obtained from Imgenex (San Diego, CA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Molecular Biology International

Article Title: Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4 α Isoform in Humans

doi: 10.1155/2016/3105478

Figure Lengend Snippet: Western blot analysis of protein extracts from human adult and fetal hearts with CH1 and TPM311 antitropomyosin antibodies. (a) Ponceau-stained blot: lane 1: extract of adult heart; lane 2: extract of fetal heart; lane M: molecular weight markers. (b) Blots stained with CH1 monoclonal antibodies that recognize all sarcomeric TPM isoforms as well as TPM4 δ with exon 9a C-terminus end. (c) Blot stained with TM311 monoclonal antibodies that recognize all high molecular wt TPM isoforms with exon 1a at the N-terminus end. However, it does not recognize TPM4 δ with exon 1b at the N-terminus end.

Article Snippet: Human adult and fetal cardiac tissue RNA were obtained from Zyagen (San Diego, CA); adult human heart protein samples were obtained from Imgenex (San Diego, CA).

Techniques: Western Blot, Staining, Molecular Weight, Bioprocessing