Twist-1 Antibody (927403) [Unconjugated]

Western blot shows lysates of PC-3 human prostate cancer cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Twist-1 Monoclonal Antibody (Catalog # MAB6230) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Twist-1 at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human peripheral blood mononuclear cell (PBMCs) were stained with Mouse Anti-Human Twist-1 Monoclonal Antibody (Catalog # MAB6230, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

DS regulated the adhesion of the aggregates of the H9 cells via canonical Wnt signaling. A GSEA analyses of Wnt-associated genes enriched in the aggregates of the H9 cells treated with DS. B Gene expression analysis by qRT-PCR for Wnt-associated genes, relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. C Immunofluorescence analysis of beta -catenin (red) on the aggregates of the H9 cells treated with or without DS; the nuclei were stained with DAPI. Scale bars = 50 μm. D The protein levels of total beta -catenin in nuclei and cytoplasm were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; the purity of nuclear and cytosolic fractions was verified using laminB1 and GAPDH, respectively. E Gene expression analysis by qRT-PCR for Wnt-targeted genes; relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. F The protein levels of WNT7B, FZD8, total beta -catenin, LEF1, MMP3, MMP7, TWIST1, SNAI2, and E-cad were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; GAPDH was used as a loading control. Data represent the mean ± SD. *P < 0.05 and **P < 0.01 and ***P < 0.001. FZD, frizzled class receptor; LEF1, lymphoid enhancer-binding factor 1; TWIST, twist family bHLH transcription factor; SNAI, snail family transcriptional repressor; MMP, matrix metallopeptidase Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35619172), licensed under a CC-BY license. Not internally tested by R&D Systems.

DS regulated the adhesion of the aggregates of the H9 cells via canonical Wnt signaling. A GSEA analyses of Wnt-associated genes enriched in the aggregates of the H9 cells treated with DS. B Gene expression analysis by qRT-PCR for Wnt-associated genes, relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. C Immunofluorescence analysis of beta -catenin (red) on the aggregates of the H9 cells treated with or without DS; the nuclei were stained with DAPI. Scale bars = 50 μm. D The protein levels of total beta -catenin in nuclei and cytoplasm were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; the purity of nuclear and cytosolic fractions was verified using laminB1 and GAPDH, respectively. E Gene expression analysis by qRT-PCR for Wnt-targeted genes; relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. F The protein levels of WNT7B, FZD8, total beta -catenin, LEF1, MMP3, MMP7, TWIST1, SNAI2, and E-cad were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; GAPDH was used as a loading control. Data represent the mean ± SD. *P < 0.05 and **P < 0.01 and ***P < 0.001. FZD, frizzled class receptor; LEF1, lymphoid enhancer-binding factor 1; TWIST, twist family bHLH transcription factor; SNAI, snail family transcriptional repressor; MMP, matrix metallopeptidase Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35619172), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow
• Clone: 927403
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Twist-1 Antibody (927403) [Unconjugated] Summary

• Immunogen: E.coli-derived recombinant human Twist-1
  Met1-His202
  Accession # Q15672
• Specificity: Detects human Twist-1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: TWIST1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Twist-1 Antibody (927403) [Unconjugated]

• acrocephalosyndactyly 3
• ACS3
• B-HLH DNA binding protein
• BHLHA38
• bHLHa38BPES3
• BPES3
• Class A basic helix-loop-helix protein 38
• CRS1
• H-twist
• H-twistblepharophimosis, epicanthus inversus and ptosis 3
• SCSBPES2
• twist homolog 1 (Drosophila)
• TWIST homolog of drosophila
• TWIST
• Twist1
• Twist-1
• twist-related protein 1

Background

Twist-1 (Twist-related protein 1; also H-Twist and bHLHa38) is a 25‑27 kDa class B member of the bHLH transcription factor family of proteins. It is widely expressed in embryo, and select adult cells such as white adipocytes. In fat, Twist-1 induces fatty acid oxidation via PGC-1 alpha and CPT-1, and promotes MCP-1 and TNF-alpha secretion by adipocytes. In epithelium, Twist-1 dysregulation represses E-Cadherin and induces N-Cadherin expression, resulting in an epithelial-to-mesenchymal transition that can lead to cancer. Human Twist-1 is 202 amino acids (aa) in length. It contains a bHLH domain (aa 109-164) with an embedded DNA-binding motif. Twist-1 forms homodimers, and heterodimerizes with TCF3, HAND 1 and HAND 2. Two distinct mutations exist that may impact its dimerization pattern. There is one seven aa insertion after Ile135, and a second seven aa insertion after Pro139. A third unrelated variant shows a four Gly insert after Gly92. Full-length human Twist-1 shares 98% aa identity with mouse Twist-1.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Twist-1 Antibody (MAB6230)(1)

Publication using MAB6230

• Rachel L. Clifford, Chen Xi Yang, Nick Fishbane, Jamie Patel, Julia L. MacIsaac, Lisa M. McEwen et al. TWIST1 DNA methylation is a cell marker of airway and parenchymal lung fibroblasts that are differentially methylated in asthma Clinical Epigenetics 2020-12-01 [PMID: 33008450] (Western Blot, Human)

Bioinformatics

• Gene Symbol: TWIST1
• Uniprot:
  • Q15672()