clptm1l Antibody

What is CLPTM1L and what alternative names are used in the literature?

CLPTM1L (cleft lip and palate transmembrane protein 1-like protein) is also known as CRR9 (cisplatin resistance-related protein 9). This protein is approximately 62.2 kilodaltons in mass and has been identified as overexpressed in various tumor types, particularly those exhibiting resistance to cisplatin. The gene encodes a transmembrane protein that shares structural similarities with the cleft lip and palate transmembrane protein family .

Where is CLPTM1L primarily localized within cells?

CLPTM1L is predominantly localized in the cytoplasm, with particularly dense expression in the perinuclear region. Immunofluorescence studies have confirmed this distribution pattern. Some research indicates that CLPTM1L may specifically associate with the endoplasmic reticulum, which aligns with its putative functions in protein processing pathways. There are also reports of potential localization to the mitochondria and plasma membrane in certain cell types .

What orthologs of CLPTM1L have been identified across species?

Based on gene homology, CLPTM1L orthologs have been identified in several mammalian species including canine, porcine, monkey, mouse, and rat models. This conservation across species suggests evolutionary importance and provides multiple model systems for investigating CLPTM1L function .

Which cancer types show significant CLPTM1L overexpression?

CLPTM1L has been found to be highly expressed in multiple cancer types including oral squamous cell carcinoma (OSCC), cervical cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, glioma, melanoma, and basal cell carcinoma. This widespread overexpression across diverse cancer types suggests a fundamental role in tumorigenesis rather than tissue-specific functions .

What is known about the role of CLPTM1L in chemoresistance?

CLPTM1L was originally identified as an overexpressed protein in cisplatin-resistant human ovarian tumor cells. The functional relationship between CLPTM1L and chemoresistance has been further validated in experimental models. In cervical cancer cell lines, siRNA-mediated knockdown of CLPTM1L enhanced sensitivity to cisplatin, suggesting that CLPTM1L may protect cancer cells from cisplatin-induced apoptosis .

What criteria should researchers consider when selecting a CLPTM1L antibody?

When selecting a CLPTM1L antibody, researchers should consider:

• Target region specificity (N-terminal, middle region, or C-terminal antibodies are available)

• Validated applications (WB, IHC, IP, ICC, IF, ELISA)

• Species reactivity (human-specific vs. cross-reactive with other species)

• Clonality (monoclonal for specific epitopes vs. polyclonal for broader detection)

• Purification method (affinity-purified antibodies typically offer higher specificity)

• Literature validation (previously published work demonstrating specificity and utility)

The commercial landscape currently includes at least 106 CLPTM1L antibodies across 20 suppliers, providing multiple options for different experimental needs .

How can researchers validate CLPTM1L antibody specificity?

A comprehensive validation approach should include:

• Positive and negative control tissues with known CLPTM1L expression levels

• siRNA/shRNA knockdown experiments to confirm signal reduction

• Peptide competition assays to verify epitope specificity

• Western blotting to confirm the detection of a single band at the expected molecular weight (approximately 62.2 kDa)

• Comparison of subcellular localization patterns with published data (perinuclear cytoplasmic localization with potential ER association)

• Cross-validation with multiple antibodies targeting different epitopes

What are the optimal methods for CLPTM1L detection in tissue samples?

For immunohistochemical detection of CLPTM1L in tissue samples:

• Formalin-fixed, paraffin-embedded sections are suitable for CLPTM1L detection

• Antigen retrieval methods should be optimized based on the specific antibody requirements

• A scoring system combining staining intensity and percentage of positive cells should be employed for quantitative assessment

• Weighted scoring approaches (multiplying intensity by percentage) provide more nuanced assessment

• A cut-off value determination (such as the value of 6 used in cervical cancer studies) using ROC curve analysis can help stratify expression levels for correlation with clinical outcomes

How should researchers approach CLPTM1L immunofluorescence experiments?

For optimal immunofluorescence detection:

• Use cytoskeletal markers (e.g., Actin) as internal references to provide context for cellular morphology

• Focus on perinuclear cytoplasmic regions where CLPTM1L is most abundant

• Consider co-staining with ER markers to evaluate association with this organelle

• Employ confocal microscopy for precise subcellular localization assessment

• Include appropriate negative controls using CLPTM1L knockdown cells

What techniques are effective for studying CLPTM1L function in cancer cells?

Functional studies of CLPTM1L can be approached through:

• siRNA-mediated knockdown to reduce CLPTM1L expression

• Verification of knockdown efficiency using qRT-PCR and western blotting

• Phenotypic assays to assess cancer hallmarks:

  • Cell proliferation (e.g., CCK-8 assay)

  • Cell migration (e.g., scratch-wound test)

  • Cell invasion (e.g., transwell assay)

• Chemosensitivity assays with cisplatin or other drugs following CLPTM1L knockdown

• Analysis of related signaling pathways based on functional enrichment analyses

How does CLPTM1L contribute to cancer progression at the molecular level?

Based on bioinformatic analyses and experimental evidence, CLPTM1L appears to be involved in several cellular processes relevant to cancer:

• Protein processing in the endoplasmic reticulum

• Protein folding

• Endoplasmic reticulum formation

• N-glycan biosynthesis

• Protein hydroxylation

These functions align with its subcellular localization and suggest roles in proteostasis and post-translational modifications that may support cancer cell survival and proliferation .

What protein interactions have been identified for CLPTM1L?

String database analysis has identified several potential interacting partners for CLPTM1L including:

• TERT (telomerase reverse transcriptase) - the catalytic subunit of telomerase that maintains telomere length and chromosome stability

• SLC6A18

• LPCAT1

• ATF7IP

The interaction with TERT is particularly significant given the role of telomerase in cancer cell immortalization .

How can CLPTM1L expression be leveraged as a biomarker in clinical settings?

CLPTM1L expression has significant potential as a biomarker in multiple contexts:

For clinical application, standardized IHC protocols with validated scoring systems (such as the weighted score with a cut-off value of 6) can stratify patients into risk groups. In cervical cancer, this approach achieved 73.3% sensitivity and 80.0% specificity for predicting recurrence .

What are the key considerations when interpreting CLPTM1L expression data across different studies?

When comparing CLPTM1L expression data across studies, researchers should consider:

• Antibody differences (epitope recognition, clonality, sensitivity)

• Scoring methodologies (H-score vs. weighted score vs. percentage positive)

• Cut-off values for defining "high" vs. "low" expression

• Tissue-specific expression patterns

• Patient cohort characteristics (cancer type, stage, treatment history)

• Correlation with other molecular markers and clinical parameters

How should researchers address potential contradictions in CLPTM1L functional studies?

To reconcile contradictory findings:

• Carefully evaluate cell line models used (different cancer types may show different CLPTM1L functions)

• Consider knockdown efficiency and potential compensatory mechanisms

• Evaluate experimental timelines (acute vs. chronic CLPTM1L depletion)

• Assess the broader molecular context through pathway analyses

• Integrate findings with patient data to determine clinical relevance

• Repeat critical experiments with multiple methodological approaches

What are emerging areas of CLPTM1L research beyond current applications?

Promising future directions include:

• Development of CLPTM1L-targeted therapies to overcome chemoresistance

• Investigation of CLPTM1L in combination with other prognostic markers for improved risk stratification

• Exploration of CLPTM1L's role in additional cancer types

• Deeper mechanistic studies of CLPTM1L's interactions with telomerase and implications for cellular immortalization

• Evaluation of CLPTM1L as a blood-based biomarker through liquid biopsy approaches

What technological advances might enhance CLPTM1L-focused research?

Emerging technologies with potential application to CLPTM1L research include:

• CRISPR-Cas9 genome editing for more precise functional studies

• Single-cell analysis to evaluate heterogeneity of CLPTM1L expression within tumors

• Spatial transcriptomics to map CLPTM1L expression patterns within the tumor microenvironment

• Proteomics approaches to comprehensively identify CLPTM1L binding partners

• Structural biology techniques to elucidate the protein's functional domains and potential druggable sites