tmem201 Antibody

What is TMEM201 and why is it important in cellular research?

TMEM201 (Transmembrane Protein 201) is an inner nuclear membrane protein with a canonical length of 666 amino acid residues and a mass of 72.2 kDa in humans. It is also known as spindle-associated membrane protein 1 (SAMP1) or NET5. TMEM201 has significant research importance because:

• It regulates nuclear movement during fibroblast polarization and migration

• It maintains endothelial cell function and angiogenesis

• It interacts with the linker of nucleoskeleton and cytoskeleton (LINC) complex

• It is expressed ubiquitously across tissues, with notable expression in colon and duodenum

Understanding TMEM201 function provides critical insights into cellular migration, nuclear positioning, and tissue development processes.

Which applications are most suitable for TMEM201 antibody detection?

Based on validated experimental data, TMEM201 antibodies are most effectively used in:

Western blotting represents the gold standard for TMEM201 detection, with observed molecular weights typically ranging from 43-45 kDa , though the theoretical weight is 72.2 kDa .

What are the optimal storage conditions for TMEM201 antibodies?

For maximum stability and performance of TMEM201 antibodies:

• Store at -20°C for long-term preservation

• Avoid repeated freeze-thaw cycles that can degrade antibody quality

• Most commercial preparations are supplied in PBS buffer containing 50% glycerol and 0.02% sodium azide

• Antibodies remain stable for approximately one year after shipment under proper storage conditions

• Aliquoting is generally unnecessary for -20°C storage in glycerol-containing buffers

How should I optimize Western blot protocols for TMEM201 detection?

When designing Western blot experiments for TMEM201:

• Sample preparation:

  • Brain tissue has shown reliable detection in mouse models

  • Human samples from various tissues including colon and duodenum are suitable

• Recommended protocol adjustments:

  • Use 1:200-1:1000 dilution for primary antibody incubation

  • Incubate at room temperature for approximately 1.5 hours

  • Consider reducing conditions and appropriate immunoblot buffer systems (Group 1 buffers have shown efficacy)

  • Be prepared to visualize bands at 43-45 kDa, which differs from the predicted 72.2 kDa mass

• Controls:

  • Include mouse brain tissue as a positive control

  • Validate specificity through knockdown/knockout controls when possible

What are the key considerations when selecting between different TMEM201 antibody products?

Research-critical factors to evaluate include:

The N-terminal domain of TMEM201 has particular functional importance, so antibodies recognizing this region may be especially valuable for mechanistic studies .

How can TMEM201 antibodies be used to investigate nuclear envelope integrity and cell migration?

Advanced research applications include:

• Co-immunoprecipitation studies:

  • TMEM201 antibodies can be used to pull down interacting partners like SUN2 and LaminA/C to study LINC complex interactions

  • Requires careful optimization of buffer conditions and antibody concentrations

• Nuclear migration assays:

  • Combining TMEM201 antibody staining with live-cell imaging can reveal dynamics of nuclear positioning during migration

  • Depletion of TMEM201 using shRNA approaches impedes EC migration and angiogenesis, providing a model system for rescue experiments

• Mechanistic domain analysis:

  • The N-terminus of TMEM201 interacts with the LINC complex via highly conserved CxxC motifs that form zinc fingers

  • Antibodies specifically recognizing this domain can be instrumental in structure-function studies

Methodologically, researchers should combine TMEM201 antibody detection with truncation mutants to dissect the functional importance of different protein domains.

How do I address discrepancies between predicted and observed molecular weights in TMEM201 detection?

This represents a common challenge in TMEM201 research:

• The canonical protein has a predicted molecular weight of 72.2 kDa

• Many studies observe bands at 43-45 kDa

• Alternative isoforms (up to 2 reported) may explain some differences

Resolution approaches:

• Validation strategies:

  • Use multiple antibodies targeting different epitopes

  • Include knockdown/knockout controls to confirm specificity

  • Perform N-terminal and C-terminal tagging to determine processing patterns

• Technical considerations:

  • Post-translational modifications may alter migration patterns

  • Membrane proteins often migrate aberrantly on SDS-PAGE

  • Consider native versus denaturing/reducing conditions

• Analysis approach:

  • Document both observed and expected weights in publications

  • Specify the specific isoform being targeted when possible

How can I use TMEM201 antibodies to investigate angiogenesis mechanisms?

Based on recent findings that TMEM201 regulates endothelial cell migration and angiogenesis , researchers can:

• Experimental design recommendations:

  • Combine TMEM201 antibody staining with tube formation assays

  • Use fibrin gel bead sprouting assays with TMEM201 antibody co-staining

  • Implement wound healing assays with TMEM201 knockdown/overexpression

• Technical approach:

  • TMEM201 depletion via shRNA significantly impedes HUVEC angiogenic behavior

  • The N-terminal region is required for proper function

  • Quantify tube formation and migration rates as functional readouts

• Mechanistic investigations:

  • Assess co-localization of TMEM201 with SUN2 and LaminA/C using confocal microscopy

  • Perform co-immunoprecipitation experiments to determine interaction dependencies

  • Employ truncation mutants to map functional domains

The research revealed TMEM201-LINC complex interactions are critical for endothelial cell migration, providing a foundation for angiogenesis-focused experiments .

What are the methodological considerations for TMEM201 antibody use in cancer research models?

Recent evidence suggests TMEM201 may modulate tumor development:

• TMEM201 positively regulates invasion and migration of breast cancer cells

• TMEM201 contributes to hepatocarcinogenesis

When designing cancer research experiments:

• Model selection:

  • Human cancer cell lines expressing TMEM201

  • Patient-derived xenografts

  • Clinical tumor samples for IHC analysis

• Methodological approaches:

  • Combine TMEM201 antibody staining with invasion/migration assays

  • Assess TMEM201 expression levels in different tumor stages

  • Correlate TMEM201 levels with patient outcomes and metastatic potential

• Analytical considerations:

  • Account for heterogeneity in TMEM201 expression across tumor regions

  • Consider nuclear envelope integrity as a functional readout

  • Examine relationships between TMEM201 and established cancer markers

Emerging evidence suggests TMEM201 may represent a novel therapeutic target, particularly in contexts where cell migration drives pathogenesis.

What are common pitfalls in TMEM201 antibody experiments and how can they be resolved?

How should researchers validate TMEM201 antibody specificity in their experimental systems?

A comprehensive validation strategy includes:

• Genetic validation:

  • Use TMEM201 knockout/knockdown controls in Western blot and immunostaining

  • RNA interference using shRNA targeting TMEM201 (published sequences available)

  • CRISPR-Cas9 gene editing for complete knockout

• Technical validation:

  • Pre-absorption with immunizing peptide where available

  • Compare results using multiple antibodies against different epitopes

  • Include positive control tissues (brain tissue is recommended)

• Bioinformatic validation:

  • Verify TMEM201 expression in target tissues using public databases

  • Check for potential cross-reactive proteins with similar epitopes

  • Review literature for expected localization and molecular weight patterns

Thorough validation ensures reliable interpretation of experimental results and strengthens publication quality.