MYO1B Antibody

What is MYO1B and what cellular functions does it regulate?

MYO1B is a single-headed membrane-associated motor protein belonging to the class I myosin superfamily. It functions primarily as an actin-based molecular motor involved in cell migration and motility . MYO1B plays critical roles in cytoskeletal organization, membrane trafficking, and cell shape regulation. At the molecular level, MYO1B promotes F-actin rearrangement through the ROCK2/LIMK/Cofilin axis by enhancing RhoA activation, which subsequently affects focal adhesion assembly . Recent studies have demonstrated its involvement in autophagy regulation, specifically in modulating autophagosome-lysosome fusion, suggesting broader cellular functions than previously understood .

How is MYO1B expression altered in different cancer types?

MYO1B expression shows significant upregulation in several cancer types compared to normal tissues. In colorectal cancer (CRC), MYO1B expression is increased in most tumor tissues compared to adjacent normal mucosa, with higher expression positively associated with tumor incidence, metastasis, and poor prognosis . Similar patterns have been observed in Arecoline-associated oral carcinoma, where MYO1B was identified as a key gene in tumorigenesis . Immunohistochemical analysis consistently shows higher MYO1B staining scores in cancer tissues, with expression levels correlating with lymph node metastasis and unfavorable outcomes . The overexpression pattern appears consistent across multiple cancer types, suggesting a common oncogenic role for MYO1B.

What is the molecular weight of MYO1B and how do I ensure I'm detecting the correct protein?

The calculated molecular weight of MYO1B is reported as 132 kDa and 125 kDa, reflecting potential isoforms . When performing western blot analysis to detect MYO1B, researchers should verify bands at these molecular weights. To ensure specificity:

• Use positive control tissues known to express MYO1B (human heart tissue, mouse brain tissue)

• Include negative controls where MYO1B expression is minimal

• Compare your results with published literature showing MYO1B detection

• Consider using multiple antibodies targeting different epitopes of MYO1B for validation

• When possible, confirm specificity through siRNA knockdown experiments

What are the optimal protocols for MYO1B immunohistochemical detection?

Based on validated protocols for MYO1B antibody (such as 15012-1-AP), the following parameters are recommended for optimal immunohistochemical (IHC) detection :

For percent positivity scoring: 0 (0%), 1 (1-25%), 2 (26-50%), 3 (51-75%), and 4 (>75%) . For staining intensity: 0 (no staining), 1 (weak/light yellow), 2 (moderate/yellowish-brown), and 3 (strong/brown) . The final expression score is calculated as the product of these two scores, ranging from 0-12.

How can I establish MYO1B knockdown or overexpression models for functional studies?

For manipulating MYO1B expression in experimental models, the following approaches have been successfully implemented:

• MYO1B knockdown models:

  • Use lentivirus-based short hairpin RNA (shRNA) targeting MYO1B mRNA

  • Verified in HCT116 colorectal cancer cell lines, which naturally express high levels of MYO1B

  • Validation of knockdown efficiency should be performed by western blot analysis

• MYO1B overexpression models:

  • Utilize lentivirus vectors containing MYO1B oligonucleotides

  • Successfully established in SW480 cell lines, which have relatively low endogenous MYO1B expression

  • Confirm overexpression through immunoblot assays

In both cases, appropriate controls (scrambled shRNA or empty vector) must be included, and multiple clones should be tested to account for clonal variation effects.

What functional assays are most informative when studying MYO1B's role in cancer progression?

Based on published research, these assays have provided meaningful insights into MYO1B function :

• Migration assays:

  • Transwell migration assays (without matrigel)

  • Wound-healing/scratch assays to assess cell motility over time

• Invasion assays:

  • Transwell assays with matrigel coating

  • 3D matrix invasion assays for more physiologically relevant assessment

• Proliferation assays:

  • MTT/CCK-8 assays

  • Colony formation assays

  • Cell cycle analysis by flow cytometry

• Angiogenesis assays:

  • HUVEC tube formation assays using conditioned media from MYO1B-modified cells

  • Analysis of VEGF secretion through ELISA

• Autophagy assessment:

  • Monitoring LC3-I to LC3-II conversion

  • Assessment of autophagosome-lysosome fusion

  • p62/SQSTM1 degradation analysis

Each assay should include appropriate positive and negative controls, and results should be validated using multiple experimental approaches.

How can I appropriately analyze MYO1B expression data from patient cohorts?

When analyzing MYO1B expression in patient samples, consider these validated analytical approaches :

• Statistical comparisons between groups:

  • Student's t-test for comparing MYO1B levels between normal and cancer tissues

  • Chi-square tests to assess associations between MYO1B expression and clinicopathological features

  • Mann-Whitney U test for non-parametric comparisons

• Survival analysis:

  • Kaplan-Meier survival curves stratified by MYO1B expression levels

  • Log-rank tests to determine statistical significance

  • Univariate and multivariate Cox regression analyses to assess MYO1B as an independent prognostic factor

• Correlation analyses:

  • Pearson or Spearman correlation to evaluate relationships between MYO1B and other molecular markers

  • Analysis of MYO1B correlation with immune cell infiltration parameters

Data should be adjusted for potential confounding variables, and multiple cohorts should be analyzed when possible to confirm findings.

What criteria should be used to classify patients as MYO1B-high versus MYO1B-low expressors?

Based on established research methodologies, the following approaches for patient stratification are recommended :

• For IHC-based classification:

  • Calculate composite scores based on staining intensity and percentage of positive cells

  • Scores ≥6 (on a 0-12 scale) are typically classified as high expression

  • Consider tissue-specific median values as cutoffs for specific cancer types

• For mRNA expression analysis:

  • Determine median expression values across the cohort

  • Use ROC curve analysis to identify optimal cutoff values for outcome prediction

  • Consider quartile-based approaches (top 25% vs. bottom 25%) for extreme phenotype comparisons

Importantly, cutoff values should be established before outcome analysis to avoid bias, and sensitivity analyses using different thresholds should be performed to assess robustness of findings.

What is known about the relationship between MYO1B and cancer metastasis?

MYO1B plays crucial roles in promoting cancer metastasis through multiple mechanisms :

• Cytoskeletal reorganization:

  • MYO1B promotes F-actin rearrangement through the ROCK2/LIMK/Cofilin axis

  • Enhances RhoA activation, leading to improved cell motility and invasion capacity

  • Facilitates focal adhesion assembly, critical for cellular movement during metastasis

• Clinical correlations:

  • High MYO1B expression positively correlates with lymph node metastasis in both oral cancer and CRC

  • Associated with distant metastasis in CRC patients

  • Independent predictor of poor prognosis, likely due to its pro-metastatic functions

• Experimental evidence:

  • MYO1B silencing significantly inhibits migration and invasion of cancer cells in vitro

  • Suppression of MYO1B reduces metastatic potential in vivo in animal models

  • Functional studies demonstrate its importance for both Arecoline-transformed oral cells and established cancer cell lines

These findings collectively establish MYO1B as a critical regulator of the metastatic process across multiple cancer types.

How does MYO1B influence tumor angiogenesis and what mechanisms are involved?

Recent research has revealed MYO1B as a significant regulator of tumor angiogenesis through the following mechanisms :

• Autophagy inhibition:

  • MYO1B inhibits autophagosome-lysosome fusion, a critical step in the autophagic process

  • This inhibition prevents the autophagic degradation of HIF-1α (Hypoxia-Inducible Factor 1α)

  • Accumulated HIF-1α subsequently enhances VEGF secretion from cancer cells

• VEGF secretion enhancement:

  • MYO1B overexpression leads to increased VEGF secretion from colorectal cancer cells

  • Elevated VEGF promotes endothelial cell recruitment and new vessel formation

  • This creates a favorable microenvironment for tumor growth and metastasis

• Microenvironment modulation:

  • MYO1B expression positively correlates with infiltration of macrophages, B cells, and dendritic cells

  • This immune cell infiltration pattern may further promote angiogenesis through additional cytokine production

This multilayered influence on angiogenesis positions MYO1B as a potential target for anti-angiogenic therapeutic strategies in cancer.

What signaling pathways interact with MYO1B in cancer progression?

MYO1B intersects with several critical signaling pathways in cancer :

• RhoA/ROCK signaling axis:

  • MYO1B enhances RhoA activation

  • Activated RhoA stimulates ROCK2, leading to LIMK/Cofilin phosphorylation

  • This cascade regulates cytoskeletal dynamics critical for cell movement and invasion

• Wnt signaling pathway:

  • MYO1B shows a close relationship with SMAD3, which is enriched in the Wnt signaling pathway

  • Wnt pathway dysregulation is fundamental to multiple cancer types, particularly colorectal cancer

• HIF-1α/VEGF pathway:

  • MYO1B stabilizes HIF-1α by inhibiting its autophagic degradation

  • Stabilized HIF-1α drives VEGF expression and secretion

  • This promotes angiogenesis and creates a hypoxic microenvironment favorable for tumor growth

• Autophagy regulation:

  • MYO1B interferes with autophagosome-lysosome fusion

  • This disruption affects multiple downstream cellular processes including protein degradation and metabolism

  • May influence cancer cell survival under stress conditions

Understanding these pathway intersections provides opportunities for combination therapeutic approaches targeting MYO1B and its interacting partners.

What are common challenges when using MYO1B antibodies and how can they be addressed?

Researchers often encounter these challenges when working with MYO1B antibodies :

How can I optimize MYO1B detection for specific tissue types?

Different tissues may require specific optimization strategies for optimal MYO1B detection :

• For colorectal tissues:

  • Paraffin-embedded tissues show good results with the LSAB protocol

  • Score both intensity and percent positivity separately before calculating composite scores

  • Compare tumor regions with adjacent normal mucosa within the same slides when possible

• For oral cancer tissues:

  • Consider relationship with Arecoline exposure in experimental design

  • MYO1B shows associations with lymph node metastasis and unfavorable outcomes

• For other tissue types:

  • Begin with recommended dilution ranges (1:20-1:200) and adjust based on pilot experiments

  • Include known positive controls (heart, brain) alongside your experimental tissues

  • Consider double-staining approaches to evaluate co-localization with interacting proteins

Regardless of tissue type, titration of antibody concentration is essential for optimal results, and all protocols should be validated for your specific experimental conditions.