albA Antibody

What is the ALBA protein and what cellular functions does it participate in?

The ALBA protein serves as a key player in DNA packaging and repair processes. It functions as a chromatin-binding protein that is instrumental in maintaining genome stability and regulating gene expression. Its involvement in DNA repair and chromosome organization highlights its significance in cellular processes . The protein is distributed uniformly and abundantly on chromosomes, suggesting a critical role in chromatin architecture. Understanding ALBA's function is essential for advancing knowledge in genetic disorders, cancer research, and developmental biology .

What validated applications are available for albA antibody in research settings?

The ALBA Polyclonal Antibody (PACO50846) has been validated for multiple research applications, primarily Western blot (WB) and ELISA techniques. Testing demonstrates its effectiveness at dilutions of 1:500-1:5000 for Western blot applications and 1:2000-1:10000 for ELISA procedures . The antibody has been confirmed to react with Pyrococcus furiosus samples and successfully detects recombinant protein in Western blot applications with an observed band size of 27 kDa, matching the predicted molecular weight .

How does the albA antibody compare to other antibodies targeting DNA-binding proteins?

The albA antibody specifically targets the ALBA protein, which binds double-stranded DNA tightly but without sequence specificity . This distinguishes it from sequence-specific DNA-binding protein antibodies used in transcription factor research. While many chromatin-associated protein antibodies require cross-linking fixation for effective immunoprecipitation, the abundant chromosomal distribution of ALBA may provide advantages in chromatin architecture studies. The polyclonal nature of PACO50846 offers broad epitope recognition, potentially providing more robust detection than single-epitope monoclonal alternatives.

What are the critical parameters for optimizing albA antibody in Western blot applications?

For optimal Western blot results using albA antibody (PACO50846), researchers should adhere to these critical parameters:

• Dilution: Use within the validated range of 1:500-1:5000, with optimal results typically achieved at 3.4μg/ml .

• Secondary antibody: Pair with goat polyclonal to rabbit IgG at 1/50000 dilution for specific detection .

• Sample preparation: Ensure complete protein denaturation and use fresh samples when possible.

• Blocking: Use 5% non-fat dry milk or BSA in TBST to minimize background.

• Incubation time: Optimize based on signal strength, typically 1-2 hours at room temperature or overnight at 4°C.

• Controls: Include positive controls (recombinant ALBA protein) and negative controls to validate specificity.

What is the recommended procedure for antibody elution using the ALBA Elution Kit?

The ALBA Elution Kit provides a standardized method for acid elution of antibodies from intact red blood cells. The recommended procedure follows these steps:

• Wash an aliquot of red blood cells once in isotonic saline, using sufficient cells to yield 1 mL of packed cells .

• Perform four washes with Working Wash Solution to remove unbound antibodies, reserving a small aliquot of the final wash supernatant as a control .

• Transfer 1 mL of washed red blood cells to a clean tube for the elution process .

• Follow the remaining protocol steps for antibody elution and neutralization.

Validation studies have demonstrated 100% equivalence in results between eluates produced by the ALBA Elution Kit and comparator kits for in vivo IgG sensitized RBCs . Repeatability testing showed 100% consistency in manual tube tests and Bio-Rad ID-System, with 80% consistency in ORTHO ID-MTS™ system .

What are the optimal storage conditions for maintaining albA antibody activity?

The ALBA Polyclonal Antibody (PACO50846) should be stored in its liquid form in the provided storage buffer containing 50% glycerol, 0.01M PBS (pH 7.4), and 0.03% Proclin 300 as a preservative . For long-term stability, storage at -20°C is recommended, with minimal freeze-thaw cycles to preserve activity. Aliquoting the antibody upon receipt can prevent repeated freeze-thaw cycles. When in use, short-term storage at 4°C (up to one week) is acceptable, but the antibody should be returned to -20°C for longer periods to maintain optimal reactivity and specificity.

How should researchers validate the specificity of albA antibody in different experimental contexts?

Validating albA antibody specificity requires a multi-faceted approach:

• Positive controls: Include recombinant ALBA protein samples with known expression levels.

• Negative controls: Test in cells/tissues known not to express the target protein.

• Peptide competition assays: Pre-incubate the antibody with purified ALBA peptide to confirm signal specificity.

• Knockdown/knockout validation: Compare signals in wildtype versus ALBA-depleted samples.

• Molecular weight verification: Confirm detection at the predicted molecular weight (27 kDa for ALBA protein) .

• Cross-reactivity assessment: Test against closely related proteins to ensure specificity.

• Multiple detection methods: Validate findings using alternative antibodies or detection techniques.

What controls should be included when using albA antibody in immunoassays?

A comprehensive control strategy for albA antibody immunoassays should include:

How can researchers troubleshoot non-specific binding when using albA antibody?

When encountering non-specific binding with albA antibody, researchers should implement the following troubleshooting strategies:

• Optimize antibody dilution: Test a range of dilutions beyond the recommended 1:500-1:5000 for Western blot applications .

• Improve blocking: Extend blocking time or try alternative blocking agents (BSA, casein, or commercial blockers).

• Increase wash stringency: Use higher detergent concentrations (0.1-0.3% Tween-20) or extend washing durations.

• Reduce antibody incubation time: Shorter incubation periods may decrease non-specific binding.

• Pre-adsorb the antibody: Incubate with non-target tissues/cells to remove cross-reactive antibodies.

• Use highly purified samples: More purified protein preparations can reduce background.

• Optimize secondary antibody: Test different dilutions or alternative secondary antibodies.

• Include competing proteins: Add 1-5% BSA or non-fat dry milk to antibody diluent.

How does ALBAclone Anti-Fyb differ from other blood grouping reagents?

ALBAclone Anti-Fyb (Human/Murine Monoclonal) Blood Grouping Reagent is specifically designed for the in vitro detection and identification of human Fyb positive red blood cells through direct agglutination . As a human/murine monoclonal antibody, it offers several advantages over conventional polyclonal alternatives:

• Increased specificity for the Fyb antigen on red blood cells

• Batch-to-batch consistency due to monoclonal production

• Reduced risk of cross-reactivity with other blood group antigens

• Standardized production that meets FDA regulatory requirements

• Validated through extensive clinical testing before approval

The reagent underwent comprehensive review processes including clinical, non-clinical, statistical, and CMC product reviews before receiving FDA approval in 2017 .

What are the methodological considerations for antibody identification (ABI) in transfusion medicine?

Antibody identification in transfusion medicine requires careful methodological considerations to ensure accurate detection of clinically significant antibodies:

• Testing frequency: Evidence suggests that the conventional 72-hour testing interval may be necessary, as extending to 14 days could miss newly developed antibodies .

• Detection methods: Integration of multiple techniques (tube, gel, solid phase) may improve sensitivity.

• Patient history: Previous transfusions or pregnancies significantly impact the likelihood of developing new antibodies .

• System specificity: Some antibody systems (e.g., Kidd) may develop more rapidly, with 6 of 13 new antibodies in one study being within this system .

• Screen result interpretation: 5 of 13 new antibody identifications showed no change in antibody screen results, highlighting the importance of complete testing regardless of screening outcomes .

• Control implementation: Last wash supernatant should be tested in parallel with eluates to confirm antibody presence .

Research indicates that approximately 0.5% of all antibody identifications performed or 0.47% of all alloimmunized patients could experience delayed identification of new alloantibody specificity if testing intervals are extended .

How can researchers design experiments to study the role of ALBA protein in DNA repair mechanisms?

Designing experiments to study ALBA's role in DNA repair requires a multifaceted approach:

• Genetic manipulation: Create ALBA knockout/knockdown cell lines using CRISPR-Cas9 or RNAi, then assess DNA repair efficiency following induced damage.

• Localization studies: Use albA antibody for immunofluorescence to track ALBA protein recruitment to DNA damage sites, with temporal resolution after damage induction.

• Protein interaction assays: Employ co-immunoprecipitation with albA antibody to identify ALBA interactions with known DNA repair proteins.

• DNA binding assays: Conduct ChIP-seq using albA antibody to map ALBA binding sites genome-wide before and after DNA damage.

• Functional rescue experiments: Reintroduce wild-type and mutant ALBA constructs into knockout models to determine critical domains.

• DNA damage sensitivity assays: Compare survival of ALBA-deficient versus control cells following exposure to DNA-damaging agents.

• Real-time recruitment studies: Use live-cell imaging with fluorescently tagged ALBA to monitor recruitment kinetics to damage sites.

What statistical approaches are recommended for analyzing quantitative data obtained using albA antibody?

When analyzing quantitative data generated using albA antibody, researchers should consider these statistical approaches:

• Normalization methods: Use reference proteins (housekeeping genes) to normalize ALBA protein expression levels across samples.

• Technical replication: Perform at least three independent experimental replicates to ensure reproducibility.

• Statistical tests:

  • Use paired t-tests for before/after comparisons within the same samples

  • Apply ANOVA for multi-group comparisons, followed by appropriate post-hoc tests

  • Consider non-parametric alternatives (Mann-Whitney, Kruskal-Wallis) if data is not normally distributed

• Multiple testing correction: Apply Bonferroni or false discovery rate corrections when performing multiple comparisons.

• Power analysis: Determine appropriate sample sizes before experiments to ensure sufficient statistical power.

• Regression analysis: Consider for correlation studies between ALBA expression and other variables.

• Data visualization: Present data with appropriate error bars (standard deviation, standard error, or confidence intervals) and individual data points when possible.

How does the performance of polyclonal versus monoclonal albA antibody compare in various applications?

The choice between polyclonal and monoclonal albA antibodies significantly impacts experimental outcomes across different applications:

The ALBA Polyclonal Antibody (PACO50846) provides advantages through recognition of multiple epitopes, making it suitable for research applications where sensitivity is paramount . For clinical applications like blood grouping reagents, monoclonal antibodies like ALBAclone Anti-Fyb offer better standardization and regulatory compliance .