Recombinant Human Transmembrane protein 151B (TMEM151B)
Product Specs
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Target Background
- Observational study of gene-disease association. (HuGE Navigator) PMID: 19773279
Q&A
FAQs for Researchers on Recombinant Human TMEM151B
Recombinant Human Transmembrane protein 151B (TMEM151B) is a lesser-studied protein with emerging roles in neurobiology and oncology. Below are research-focused FAQs addressing methodological and analytical challenges, derived from genomic, structural, and functional data.
What experimental methods are recommended for detecting TMEM151B expression in human tissues?
TMEM151B exhibits tissue-specific expression, with high levels in the brain (cerebellum, medulla) and testes . To validate this:
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RNA-seq: Prioritize brain or testis samples, using primers targeting exon-exon junctions (e.g., Chr6:44,270,450–44,307,506) .
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Western blot: Use antibodies against the C-terminal region (theoretical pI 6.72, ~61 kDa) . Include controls for transmembrane domain extraction, as TMEM151B has two transmembrane helices .
How can researchers confirm the subcellular localization of recombinant TMEM151B?
TMEM151B is membrane-associated. For localization studies:
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Immunofluorescence: Co-stain with ER markers (e.g., Calnexin), given its interaction with SREBF2, an ER membrane protein .
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Fractionation assays: Isolate membrane fractions via differential centrifugation and validate using SDS-PAGE .
What structural features of TMEM151B should inform epitope selection for antibody development?
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Low lysine/arginine content: Avoid epitopes in lysine/arginine-rich regions (lysine: 1.4%, arginine: 0.8%) .
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Transmembrane domains: Target extracellular loops (amino acids 100–150 and 300–350) for higher accessibility .
How do somatic mutations in TMEM151B influence cancer progression, and how can they be analyzed?
TMEM151B mutations are linked to colorectal and post-menopausal breast cancers . Key approaches:
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COSMIC mutation analysis: Screen for recurrent mutations (e.g., missense variants at amino acids 204 and 398) .
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Functional assays: Compare wild-type and mutant TMEM151B in migration/invasion assays using CRISPR-edited cell lines .
| Mutation (Amino Acid) | Mutation Type | Frequency in COSMIC | Associated Cancer |
|---|---|---|---|
| p.Arg204Trp | Missense | 2.1% | Colorectal |
| p.Gly398Ser | Missense | 1.7% | Breast |
What strategies resolve contradictions in TMEM151B’s role in cholesterol biosynthesis?
TMEM151B interacts with SREBF2, a cholesterol regulator . If conflicting data arise:
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Knockdown/knockout models: Use siRNA in hepatic cells and measure LDL receptor expression via qPCR.
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Co-IP/MS: Validate direct binding to SREBF2 under cholesterol-depleted conditions .
How can recombinant TMEM151B be purified given its transmembrane domains?
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Detergent screening: Test n-dodecyl-β-D-maltoside (DDM) for solubilization, followed by nickel affinity chromatography (His-tagged constructs) .
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Validation: Confirm structural integrity via circular dichroism (CD) spectroscopy and thermal shift assays .
What bioinformatics tools are critical for analyzing TMEM151B’s evolutionary conservation?
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Paralog analysis: Compare TMEM151B with TMEM151A (47.7% identity) using Clustal Omega .
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Ortholog mapping: Use Ensembl to identify conserved residues in vertebrates (e.g., mouse, zebrafish) .
How to address low yields in recombinant TMEM151B production?
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Codon optimization: Adjust rare codons (e.g., AGA/AGG arginine codons) for mammalian expression systems .
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Membrane scaffolding: Co-express with lipid chaperones (e.g., Saposin A) to stabilize folding .
