ACRV1 Antibody

What is ACRV1 and why is it important in reproductive research?

ACRV1, also known as SP-10, is an acrosomal matrix protein evolutionarily conserved among mammals. It is specifically associated with the matrix of the acrosomal vesicle in developing spermatids and the acrosomal compartment. Its tissue-specific expression pattern makes it a valuable marker for studies of spermatogenesis, fertilization, and male reproductive function .

The protein's importance stems from its role in acrosomal development and function, which is critical for sperm-egg interaction. Its strict testis-specific expression has made ACRV1 an important biomarker in reproductive studies and potentially in fertility diagnostics. Research shows consistent detection in human testis tissue and various prostate cancer cell lines including DU 145, LNCaP, and PC-3 cells .

What are the molecular characteristics of the ACRV1 protein?

ACRV1 has the following molecular characteristics:

The notable difference between calculated and observed molecular weights (28 kDa vs. 13-36 kDa) suggests the protein may undergo significant post-translational modifications or processing, which researchers should consider when analyzing Western blot results .

How do I select the appropriate ACRV1 antibody for my specific research application?

When selecting an ACRV1 antibody, consider these critical factors:

• Application compatibility: Different ACRV1 antibodies are validated for specific applications. Based on the search results, antibodies like 14040-1-AP are validated for multiple applications including WB, IHC, IF/ICC, IP, and ELISA . Match your experimental needs with validated applications listed on product pages.

• Species reactivity: ACRV1 antibodies may show different species reactivity profiles. For example, the 14040-1-AP antibody has been tested specifically with human samples but has cited reactivity with mouse and rat models . The CAC11728 antibody specifically targets human ACRV1 .

• Clonality consideration:

  • Polyclonal antibodies (like 14040-1-AP and CAC11728) offer broader epitope recognition

  • Monoclonal antibodies provide higher specificity for single epitopes

• Validation evidence: Review available validation data including Western blot images, IHC staining patterns, and published citations. For rigorous research, choose antibodies with documented validation in multiple applications .

What validation methods should I use to confirm ACRV1 antibody specificity?

To ensure experimental rigor, validate your ACRV1 antibody through multiple approaches:

• Positive and negative control tissues/cells:

  • Positive controls: Human testis tissue shows consistent ACRV1 expression

  • Cell line controls: DU 145, LNCaP, and PC-3 cells show detectable expression

  • Negative controls: Non-reproductive tissues should show minimal to no expression

• Antibody comparison validation:

  Validation Method	Approach	Expected Result
  Western blot	Compare banding pattern with predicted MW	Primary band between 13-36 kDa
  Peptide competition	Pre-incubate antibody with immunizing peptide	Signal reduction/elimination
  Knockout/knockdown	Test in ACRV1-deficient samples	Absence of specific signal

• Multi-application consistency: Confirm target detection across different techniques (WB, IHC, IF) to strengthen confidence in specificity .

What are the optimal protocols for detecting ACRV1 in Western blot applications?

For optimal Western blot detection of ACRV1, follow these evidence-based recommendations:

• Sample preparation:

  • Recommended cell lines: DU 145, LNCaP, or PC-3 cells show consistent detection

  • Tissue samples: Human testis tissue lysates are ideal positive controls

  • Lysis buffer: Standard RIPA buffer with protease inhibitors

• Antibody dilution and incubation:
  Based on validated protocols, the recommended dilution range for WB is 1:1000-1:8000 . Optimize within this range for your specific sample type:

  Sample Type	Starting Dilution	Optimization Range
  Cell lines	1:2000	1:1000-1:4000
  Tissue lysates	1:1000	1:500-1:2000

• Expected results:

  • Molecular weight: Look for bands between 13-36 kDa (observed range), with the theoretical weight at 28 kDa

  • Multiple bands may represent different isoforms or post-translational modifications

• Troubleshooting considerations:

  • Optimize protein loading (25-50 μg per lane)

  • Consider gradient gels (4-20%) for better separation in the 13-36 kDa range

  • Extended transfer time may be necessary for complete protein transfer

What are the recommended protocols for ACRV1 immunohistochemistry in tissue samples?

For optimal immunohistochemical detection of ACRV1 in tissue samples:

• Sample preparation and antigen retrieval:

  • Fixation: 10% neutral buffered formalin is standard

  • Section thickness: 4-6 μm recommended

  • Critical step: Antigen retrieval with TE buffer pH 9.0 is specifically recommended for ACRV1; alternatively, citrate buffer pH 6.0 may be used

• Antibody dilution and detection:

  Application	Recommended Dilution	Incubation
  IHC	1:50-1:500	Overnight at 4°C

  Start with 1:100 dilution and optimize based on signal-to-noise ratio.

• Controls and validation:

  • Positive control: Human testis tissue shows specific staining

  • Negative control: Omit primary antibody or use isotype control

  • Expected pattern: Specific staining in developing spermatids and acrosomal regions

• Signal development and counterstaining:

  • DAB substrate provides good contrast

  • Hematoxylin counterstain at moderate intensity avoids obscuring specific signals

  • Mounting with permanent media preserves long-term staining

How can I optimize ACRV1 immunofluorescence staining in fixed cells?

For high-quality immunofluorescence detection of ACRV1:

• Cell preparation and fixation:

  • Validated cell model: PC-3 cells show consistent IF/ICC staining

  • Fixation method: 4% paraformaldehyde (15 minutes at room temperature)

  • Permeabilization: 0.1% Triton X-100 (10 minutes)

• Antibody concentration and staining protocol:

  Application	Recommended Dilution	Optimization Strategy
  IF/ICC	1:200-1:800	Titrate within range

  For co-localization studies, combine with other acrosomal markers (e.g., acrosin) to confirm specificity.

• Signal amplification options:

  • Tyramide signal amplification may enhance detection of low abundance protein

  • Biotinylated secondary antibodies with streptavidin-fluorophore can increase sensitivity

• Data acquisition parameters:

  • Use confocal microscopy for detailed subcellular localization

  • Z-stack imaging helps resolve acrosomal structures

  • Standardize exposure settings across experimental groups

What are common troubleshooting strategies for inconsistent ACRV1 antibody results?

When encountering inconsistent results with ACRV1 antibodies, consider these evidence-based troubleshooting approaches:

• Western blot issues:

  Problem	Potential Cause	Solution
  No bands observed	Insufficient protein	Increase loading to 50-75 μg
  Multiple unexpected bands	Cross-reactivity	Try alternative antibody (e.g., CAC11728)
  Incorrect MW bands	Post-translational modifications	Extended run time; consider gradient gels

• IHC/IF optimization:

  • Background reduction: Increase blocking time (3% BSA, 1 hour)

  • Weak signal: Test both recommended antigen retrieval methods (TE buffer pH 9.0 and citrate buffer pH 6.0)

  • Non-specific binding: Pre-absorb antibody with non-specific proteins

• Storage and handling considerations:

  • Aliquot antibodies upon receipt to avoid freeze-thaw cycles

  • Store at -20°C in recommended buffer (PBS with 0.02% sodium azide and 50% glycerol pH 7.3)

  • Follow manufacturer's stability guidelines (typically one year after shipment)

How can ACRV1 antibodies be applied in male fertility research studies?

ACRV1 antibodies provide valuable tools for fertility research applications:

• Acrosome reaction studies:

  • Track acrosomal integrity during capacitation and acrosome reaction

  • Correlate ACRV1 localization changes with fertilization potential

  • Combine with calcium signaling markers for comprehensive analysis

• Sperm maturation analysis:

  • Monitor ACRV1 expression patterns during spermatogenesis

  • Compare normal versus pathological development

  • Combine with flow cytometry for quantitative assessment of sperm subpopulations

• Fertility biomarker development:

  Research Application	Methodology	Expected Outcome
  Idiopathic infertility	Compare ACRV1 patterns between fertile/infertile samples	Potential diagnostic markers
  Contraceptive development	Target ACRV1 function	Disruption of acrosomal function
  ART optimization	Monitor acrosomal integrity	Improved sperm selection

• Experimental design recommendations:

  • Include age-matched controls

  • Standardize sample collection and processing

  • Combine with functional assays (e.g., zona binding)

What methodological considerations are important when using ACRV1 antibodies in co-immunoprecipitation experiments?

For successful co-immunoprecipitation (co-IP) studies with ACRV1 antibodies:

• Validated protocol parameters:

  Parameter	Recommended Value	Notes
  Antibody amount	0.5-4.0 μg	Per 1.0-3.0 mg total protein
  Validated cell type	DU 145 cells	Positive IP detection
  Lysis buffer	Non-denaturing	Preserve protein-protein interactions

• Experimental design considerations:

  • Pre-clear lysates with protein A/G beads to reduce non-specific binding

  • Include IgG control to identify non-specific interactions

  • Consider crosslinking antibody to beads for cleaner results

• Detection of interaction partners:

  • Validate interactions through reciprocal co-IP

  • Confirm with alternative methods (e.g., proximity ligation assay)

  • Consider mass spectrometry for unbiased interaction profiling

• Results interpretation:

  • Expected ACRV1 band at 13-36 kDa range

  • Focus on reproductive biology-relevant interaction partners

  • Compare interaction profiles between normal and pathological states

How can multi-color flow cytometry with ACRV1 antibodies advance sperm function research?

Multi-color flow cytometry with ACRV1 antibodies offers powerful analytical capabilities:

• Panel design for comprehensive sperm analysis:

  Marker	Purpose	Combined Interpretation
  ACRV1	Acrosomal integrity	Core marker for acrosomal status
  Annexin V	Membrane integrity	Distinguish viable from non-viable sperm
  Mitochondrial dyes	Mitochondrial function	Assess energy production capacity
  DNA fragmentation markers	Genetic integrity	Complete fertility potential profile

• Protocol optimization:

  • Fixation/permeabilization must preserve both acrosomal and membrane integrity

  • Validated ACRV1 antibody dilution for flow cytometry: typically in the 1:100-1:500 range

  • Sequential staining may be necessary for complex panels

• Data analysis approach:

  • Hierarchical gating strategy starting with viable cells

  • Correlation of ACRV1 signal intensity with functional parameters

  • Machine learning algorithms for pattern recognition in heterogeneous populations

• Research applications:

  • High-throughput screening of environmental toxicants on acrosomal function

  • Personalized fertility assessment based on multidimensional sperm parameters

  • Drug development targeting specific sperm subpopulations

What are the considerations for using ACRV1 antibodies in cross-species comparative reproductive studies?

When extending ACRV1 research across species:

• Species reactivity validation:

  • Confirmed reactivity: Human samples (tested)

  • Cited reactivity: Mouse and rat models (cited but requires validation)

  • Predicted reactivity: Based on sequence homology across mammalian species

• Cross-species experimental design:

  Species	Epitope Conservation	Validation Method
  Human	Reference standard	Direct testing
  Mouse/Rat	Moderate to high	Western blot with tissue-specific controls
  Non-human primates	High	Start with human-validated antibodies
  Other mammals	Variable	Sequence alignment before testing

• Optimization strategies:

  • Adjust antibody concentration based on epitope conservation

  • Modify incubation conditions (time/temperature) for different species

  • Species-specific blocking reagents to reduce background

• Interpretation challenges:

  • Account for species differences in acrosomal development timing

  • Consider evolutionary differences in acrosomal protein function

  • Validate findings through multiple methodological approaches