ANAPC1 Antibody

What is ANAPC1 and what is its biological function?

ANAPC1 (Anaphase Promoting Complex Subunit 1) is a subunit of the anaphase-promoting complex, an E3 ubiquitin ligase that regulates progression through the metaphase to anaphase transition during cell cycle. This complex functions by ubiquitinating specific proteins, targeting them for degradation, which is essential for proper cell cycle progression . ANAPC1 is also known by several synonyms including TSG24, APC1, and MCPR . The protein has a calculated molecular weight of 217 kDa (1944 amino acids), though it typically appears at 200-210 kDa in experimental conditions .

What types of ANAPC1 antibodies are commercially available for research?

Several types of ANAPC1 antibodies are available for research applications:

• Rabbit polyclonal unconjugated antibodies (e.g., 21748-1-AP) suitable for Western Blot, IHC, and ELISA applications

• Rabbit polyclonal antibodies specific for IHC applications (e.g., A45514)

• Rabbit polyclonal antibodies with fluorescent conjugates such as Cy5.5® for immunofluorescence applications

The selection of an appropriate antibody should be based on your experimental design, target application, and species of interest.

What are the recommended storage conditions for ANAPC1 antibodies?

Storage conditions vary slightly between products but generally follow these guidelines:

Most ANAPC1 antibodies are supplied in a stabilizing buffer containing glycerol (typically 50%) and a preservative such as sodium azide to maintain antibody integrity during storage .

What are the optimal dilutions for ANAPC1 antibodies in different applications?

Optimal dilutions for ANAPC1 antibodies vary by application type:

It is strongly recommended to titrate the antibody in each testing system to obtain optimal results as the dilution may be sample-dependent .

What antigen retrieval methods are recommended for IHC with ANAPC1 antibodies?

For immunohistochemical applications using ANAPC1 antibodies:

• Primary recommendation: Antigen retrieval with TE buffer pH 9.0

• Alternative method: Antigen retrieval with citrate buffer pH 6.0

The choice between these methods may depend on the specific tissue being examined and should be optimized for your particular experimental setup. Positive IHC detection has been reported in mouse testis tissue, human cervical cancer tissue, and human stomach tissue using these retrieval methods .

How can I validate the specificity of ANAPC1 antibodies in my experiments?

To validate ANAPC1 antibody specificity:

• Positive controls: Use cell lines with confirmed ANAPC1 expression such as HeLa cells, HEK-293 cells, HT-1080 cells, or K-562 cells

• Tissue controls: Mouse brain tissue has shown positive WB results, while mouse testis tissue, human cervical cancer tissue, and human stomach tissue have shown positive IHC results

• Overexpression validation: Compare results with transfected HEK-293 cells overexpressing ANAPC1

• Molecular weight verification: Confirm detection at the expected molecular weight of 200-210 kDa

• CRISPR knockdown: Consider using CRISPR techniques to create ANAPC1 knockdown models as negative controls, similar to approaches mentioned in LUSC research

What is known about ANAPC1 expression in cancer, particularly in lung squamous cell carcinoma (LUSC)?

Recent research has revealed significant insights into ANAPC1 expression in LUSC:

• ANAPC1 mRNA is significantly upregulated in LUSC tissues (SMD = 1.97, 95% CI [1.26–2.67])

• Protein-level analysis has confirmed this upregulation (p < 0.001)

• Higher ANAPC1 expression correlates with poorer survival in LUSC patients (HR = 1.11, 95% CI: 1–1.49)

• Expression patterns show demographic and clinical associations:

  • Higher in males compared to females

  • Higher in N1-stage compared to N0-stage

  • Lower in grade I compared to grades II/III

These findings suggest ANAPC1 may serve as a potential biomarker for LUSC progression and prognosis.

How does ANAPC1 expression influence immune cell infiltration and therapy response?

Research indicates that ANAPC1 expression levels may impact immune responses and therapy effectiveness:

• Overexpression of ANAPC1 has been associated with reduced immune cell infiltration in tumor microenvironments

• Higher ANAPC1 expression correlates with decreased effectiveness of immunotherapy approaches

• ANAPC1 knockdown experiments have demonstrated inhibition of cancer cell proliferation, suggesting its potential role in tumor growth regulation

These findings have important implications for understanding therapy resistance mechanisms and developing strategies to improve immunotherapy outcomes in patients with high ANAPC1 expression.

What pathways and co-expressed genes are associated with ANAPC1?

Analysis of high-expression co-expressed genes (HECEGs) associated with ANAPC1 has revealed:

• Most HECEGs are enriched in cell cycle-related pathways

• ANAPC1 likely participates in cell growth-related pathways that contribute to cancer advancement and progression

Understanding these molecular relationships provides insights into the mechanistic role of ANAPC1 in cellular processes and disease progression.

How can CRISPR-based techniques be utilized to study ANAPC1 function?

CRISPR techniques have been employed to validate ANAPC1's role in cancer:

• CRISPR-mediated knockdown of ANAPC1 has been shown to inhibit cell proliferation in cancer models

• This approach allows for precise investigation of ANAPC1's functional role in various cellular processes

• CRISPR validation can complement antibody-based detection methods to provide comprehensive understanding of ANAPC1 biology

Researchers studying ANAPC1 should consider incorporating CRISPR-based functional validation to strengthen their findings.

What are common challenges when using ANAPC1 antibodies and how can they be addressed?

When working with ANAPC1 antibodies, researchers may encounter several challenges:

• High molecular weight detection: At 200-210 kDa, ANAPC1 requires special consideration for efficient transfer in Western blotting:

  • Use lower percentage gels (6-8%)

  • Extend transfer time

  • Consider using PVDF membranes with larger pore sizes for better transfer of high molecular weight proteins

• Background signal: To reduce non-specific binding:

  • Optimize blocking conditions (5% non-fat milk or BSA)

  • Increase washing steps between antibody incubations

  • Consider using more stringent washing buffers (higher salt concentration)

• Variability between tissue types: Different tissues may require optimization of:

  • Fixation protocols

  • Antigen retrieval methods (TE buffer pH 9.0 or citrate buffer pH 6.0)

  • Antibody concentration (1:50-1:500 for IHC applications)

How can I select the appropriate secondary antibodies for ANAPC1 detection?

For optimal detection of ANAPC1 using rabbit polyclonal primary antibodies, consider these secondary antibody options:

• For colorimetric detection: Goat Anti-Rabbit IgG H&L Antibody (AP)

• For chemiluminescent detection: Goat Anti-Rabbit IgG H&L Antibody (HRP)

• For fluorescent detection: Goat Anti-Rabbit IgG H&L Antibody (FITC)

• For signal amplification: Goat Anti-Rabbit IgG H&L Antibody (Biotin)

The choice of secondary antibody should align with your detection system and experimental requirements.

What potential therapeutic targets are associated with ANAPC1 in cancer?

Drug sensitivity and molecular docking analyses have identified several compounds with potential for targeting ANAPC1 in cancer therapy:

• Tenovin-1: Shows binding affinity to ANAPC1 and potential antitumor activity

• Carboxyatractyloside: Identified as a potential ANAPC1-targeting therapeutic agent

• Phycocyanobilin: Demonstrated potential as an antitumor agent targeting ANAPC1

These findings suggest possible avenues for developing targeted therapies against ANAPC1-overexpressing tumors, particularly in LUSC.

How might ANAPC1 expression data be utilized for patient stratification and personalized medicine?

Based on current research findings, ANAPC1 expression patterns could inform clinical decision-making in several ways:

• Prognostic stratification: Higher ANAPC1 expression correlates with poorer survival in LUSC patients (HR = 1.11, 95% CI: 1–1.49), suggesting its potential use as a prognostic biomarker

• Treatment selection: Patients with lower ANAPC1 expression may respond better to immunotherapy approaches, informing treatment selection decisions

• Demographic considerations: The observed differences in expression between males/females and different tumor grades/stages suggest potential for tailored therapeutic approaches based on ANAPC1 expression patterns