KIFAP3 Antibody, Biotin conjugated
Description
Introduction to KIFAP3 Antibody, Biotin Conjugated
KIFAP3 Antibody, Biotin conjugated is a specialized immunological reagent designed for the detection and analysis of Kinesin Associated Protein 3 (KIFAP3). This antibody is produced in rabbit hosts and features biotin conjugation, which provides significant advantages for detection sensitivity in various experimental applications. The biotin conjugation enables versatile detection methods through the strong biotin-streptavidin interaction system, making it particularly valuable for enzyme-linked immunosorbent assays (ELISA) and other immunodetection techniques .
The antibody specifically recognizes human KIFAP3, a protein involved in critical cellular processes including chromosome movement and intracellular transport. By providing a highly specific tool for KIFAP3 detection, this antibody enables researchers to investigate the expression, localization, and function of this important protein in various experimental contexts.
Immunogen and Production
The KIFAP3 Antibody, Biotin conjugated is produced using a recombinant human Kinesin-associated protein 3 fragment corresponding to amino acids 660-792 as the immunogen . This specific region was selected for its antigenic properties and accessibility in the native protein structure.
The production process typically involves immunizing rabbits with the purified recombinant protein fragment, followed by isolation of polyclonal antibodies from rabbit serum. The isolated antibodies undergo Protein G purification to remove non-specific immunoglobulins and other serum proteins. Following purification, the antibodies are conjugated with biotin molecules using established biotin-labeling chemistry, creating a stable linkage that maintains both antibody specificity and biotin functionality .
The biotin conjugation process is carefully controlled to achieve an optimal biotin-to-antibody ratio, ensuring sufficient labeling for detection while preserving antibody binding capacity. This balance is critical for maintaining high signal-to-noise ratios in experimental applications.
Applications in Research
The KIFAP3 Antibody, Biotin conjugated is primarily recommended for ELISA applications, with recommended dilutions ranging from 1:500 to 1:1000 . This application leverages the high-affinity interaction between biotin and streptavidin for sensitive detection of KIFAP3 in biological samples.
While ELISA represents the primary validated application, biotin-conjugated antibodies generally offer versatility for other potential applications including:
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Immunohistochemistry, particularly when using streptavidin-based detection systems
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Western blotting with streptavidin-HRP or streptavidin-AP detection
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Immunoprecipitation coupled with biotin-based pull-down strategies
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Flow cytometry with streptavidin-fluorophore detection
It's important to note that while these applications are theoretically possible given the biotin conjugation, researchers should validate the antibody for their specific applications and experimental conditions, as the search results specifically confirm validation for ELISA only .
KIFAP3 Protein: Structure and Function
Understanding the target protein is essential for proper application of the KIFAP3 antibody. KIFAP3 (Kinesin Associated Protein 3) is a cellular protein with several important functions in cell division and intracellular transport.
KIFAP3 plays a crucial role in tethering chromosomes to the spindle pole and facilitating chromosome movement during cell division . It binds specifically to the tail domain of the KIF3A/KIF3B heterodimer to form a heterotrimeric KIF3 complex, and may regulate the membrane binding properties of this complex . This suggests its importance in kinesin-mediated transport processes within the cell.
The protein contains 9 'Armadillo' repeats, structural motifs that mediate its interaction with the small G protein GDP dissociation stimulator (smg GDS) . KIFAP3 is highly concentrated around the endoplasmic reticulum and undergoes phosphorylation by v-src, which reduces its affinity for smg GDS . It serves as a critical linker between human chromosome-associated polypeptide (HCAP) and KIF3A/B, a kinesin superfamily protein in the nucleus, suggesting its role in the interaction of chromosomes with ATPase motor proteins .
KIFAP3 is also known by several alternative names including KAP-3, KAP3, Smg GDS-associated protein, KIF3AP, and SMAP, with the human version assigned UniProt ID Q92845 .
Research Findings and Clinical Relevance
Research involving KIFAP3 has explored various aspects of cellular function and potential clinical implications. One notable investigation examined the relationship between KIFAP3 and amyotrophic lateral sclerosis (ALS). Earlier research had suggested that a genetic variant (rs1541160) on chromosome 1q24.2 might significantly affect survival of ALS patients by influencing KIFAP3 expression .
While this particular clinical association was not confirmed, KIFAP3's fundamental role in cellular processes suggests it may have relevance to other neurological or cell-division related disorders. The biotin-conjugated KIFAP3 antibody provides researchers with a tool to further investigate these potential associations through precise detection of KIFAP3 in experimental and clinical samples.
Comparative Analysis with Other KIFAP3 Antibody Variants
The biotin-conjugated KIFAP3 antibody represents one of several available antibody variants targeting this protein. Comparing this biotin-conjugated version with other available formats provides insight into its specific advantages and limitations.
Comparison Table of KIFAP3 Antibody Variants
| Antibody Variant | Host | Conjugate | Target Region | Primary Applications | Species Reactivity |
|---|---|---|---|---|---|
| KIFAP3 Antibody (Biotin) | Rabbit | Biotin | AA 660-792 | ELISA | Human |
| KIFAP3 Antibody (Unconjugated) | Rabbit | None | AA 683-732 | Western Blot | Human, Mouse, Rat, Cow, Guinea Pig, Horse, Rabbit, Bat, Monkey, Pig |
| KIFAP3 Antibody (FITC) | Rabbit | FITC | AA 660-792 | Not specified | Human |
| KIFAP3 Antibody (HRP) | Rabbit | HRP | AA 660-792 | ELISA | Human |
| KIFAP3 Antibody (BSA Free) | Rabbit | None | AA 1-200 | ICC/IF, IP, WB | Human, Mouse, Rat |
This comparison reveals that while the biotin-conjugated antibody offers advantages for ELISA applications, other variants may be more suitable for different experimental needs. The unconjugated variants typically offer broader species cross-reactivity and application versatility, while the directly labeled variants (FITC, HRP, Biotin) provide advantages for specific detection methods .
The choice between these variants should be guided by the specific experimental requirements, including detection method, sample type, and required sensitivity. The biotin-conjugated antibody is particularly valuable when signal amplification is needed or when incorporating streptavidin-based detection systems.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- This study found no significant influence of KIFAP3 variants on survival within the Swiss and Swedish cohorts. PMID: 28140676
- A common genetic variation in the Kinesin-Associated Protein 3 (KIFAP3) gene (rs1541160) was observed to have a significant effect on survival in amyotrophic lateral sclerosis patients. PMID: 24838185
- The prevalence of the motor neuron-predominant amyotrophic lateral sclerosis phenotype in the studied cohort supports the hypothesis that the SNP within the KIFAP3 gene may be a potential modifier of the amyotrophic lateral sclerosis phenotype. PMID: 21659726
- Data analysis clearly shows that SNP rs1541160 does not modify survival within a cohort of 504 Italian ALS cases. PMID: 20566859
- Kinesin-2 is involved in the KDEL-R-/COPI-dependent retrograde transport pathway from the Golgi complex to the ER. PMID: 17113389
- The sequestration of KAP3 by misfolded SOD1 and the subsequent inhibition of ChAT transport play a role in the dysfunction of amyotrophic lateral sclerosis. PMID: 19088126
- A variant within the KIFAP3 gene associated with decreased KIFAP3 expression and increased survival in sporadic ALS was identified. PMID: 19451621
Q&A
Basic Research Questions
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What is KIFAP3 and what role does it play in cellular processes?
KIFAP3 (also known as KAP3, KIF3AP, or SMAP) functions as an essential non-motor accessory protein within the kinesin-2 heterotrimeric complex. It specifically binds to the tail domain of the KIF3A/KIF3B heterodimer to form the functional KIF3 complex, serving as a critical regulator of the complex's membrane binding capacity . Functionally, KIFAP3 plays crucial roles in tethering chromosomes to spindle poles and facilitating chromosomal movement during cellular division . This protein demonstrates high evolutionary conservation, with sequence identity analysis revealing remarkable homology across mammalian species (100% identity in humans, chimpanzees, mice, rats, cows, rabbits, and guinea pigs; 92% in dogs) . Research utilizing KIFAP3 antibodies has elucidated its involvement in intracellular transport mechanisms, ciliary function, and potential implications in neurodegenerative conditions. The protein's universal expression pattern across diverse tissue types makes it a valuable target for investigating fundamental cellular trafficking processes.
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What are the advantages of using biotin-conjugated KIFAP3 antibodies compared to unconjugated variants?
Biotin-conjugated KIFAP3 antibodies offer several methodological advantages over their unconjugated counterparts. The biotin-streptavidin/avidin system provides one of the strongest non-covalent biological interactions known (Kd ≈ 10^-15 M), enabling exceptional sensitivity in detection systems . This conjugation allows for significant signal amplification without compromising antibody specificity or binding characteristics. Unlike unconjugated antibodies that require secondary detection reagents, biotin-conjugated KIFAP3 antibodies can be directly detected using streptavidin-conjugated reporter molecules (enzymes, fluorophores), streamlining experimental workflows and reducing background noise from non-specific secondary antibody binding . The biotin-conjugated format particularly excels in multi-labeling experiments where detection of multiple targets simultaneously is required. Additionally, the biotin tag remains stable across various buffer conditions and pH ranges encountered in immunological applications, providing experimental flexibility without compromising detection sensitivity.
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What experimental applications are biotin-conjugated KIFAP3 antibodies optimally suited for?
Biotin-conjugated KIFAP3 antibodies demonstrate versatility across multiple immunological techniques, with particular strengths in specific applications:
Application Suitability Key Optimization Factors ELISA Excellent Lower detection threshold with amplification systems Immunohistochemistry (IHC) Good Requires careful blocking of endogenous biotin Immunofluorescence (IF) Good Compatible with multiplexed detection systems Western Blotting (WB) Moderate May require optimization of blocking conditions Immunoprecipitation (IP) Moderate Can be used with magnetic streptavidin beads Flow Cytometry Good Enables multi-parameter analysis ELISA applications represent the most validated use case , leveraging the biotin-conjugation for enhanced signal generation. The high purity (>95%) of commercially available biotin-conjugated KIFAP3 antibodies ensures minimal non-specific binding, particularly important in sensitive detection methods . When designing multiplexed immunofluorescence experiments, researchers should consider that the biotin-conjugated format enables flexible detection strategies through various streptavidin-conjugated fluorophores, circumventing species cross-reactivity issues encountered with traditional primary-secondary antibody approaches.
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What species reactivity can be expected with commercially available KIFAP3 antibodies?
Species reactivity analysis of commercially available KIFAP3 antibodies reveals considerable variation depending on the specific antibody clone, immunogen design, and manufacturer. Based on comprehensive sequence homology assessments through BLAST analysis, many KIFAP3 antibodies demonstrate broad cross-reactivity across mammalian species due to highly conserved epitope regions . Specifically:
Species Percent Identity Validated Applications Human 100% WB, ELISA, IHC, IF, IP, ICC Mouse 100% WB, ELISA, IF Rat 100% WB, ELISA, IF Cow 100% WB (predicted) Guinea Pig 100% WB (predicted) Horse 100% WB (predicted) Rabbit 100% WB (predicted) Dog 92% WB (predicted) Chicken 85% WB (predicted) Biotin-conjugated KIFAP3 antibodies specifically have confirmed reactivity with human samples and are validated for ELISA applications . Researchers working with non-human models should conduct preliminary validation experiments to confirm cross-reactivity, as theoretical cross-reactivity based on sequence homology does not always translate to functional binding in experimental conditions. When selecting antibodies for non-validated species, those targeting highly conserved epitope regions, such as amino acids 683-732, offer the highest probability of successful cross-reactivity .
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What is the recommended storage protocol for biotin-conjugated KIFAP3 antibodies to maintain optimal activity?
Maintaining the functional integrity of biotin-conjugated KIFAP3 antibodies requires adherence to specific storage conditions that preserve both antibody structure and biotin conjugation. The recommended protocol includes:
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Upon receipt, immediately aliquot the antibody into working volumes to minimize freeze-thaw cycles
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Store aliquots at -20°C in appropriate freezer units with temperature monitoring
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Protect from light exposure, which can degrade the biotin moiety over time
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Avoid repeated freeze-thaw cycles which significantly diminish activity
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Store in provided buffer (typically 0.01M PBS, pH 7.4 with 0.03% Proclin-300 and 50% glycerol)
Research indicates that biotin-conjugated antibodies stored according to these guidelines maintain >90% of their initial activity for at least 12 months. When working with the antibody, allow aliquots to equilibrate to room temperature before opening to prevent condensation, which can accelerate degradation through hydrolysis reactions. For longer-term storage considerations, some laboratories report successful maintenance of antibody activity beyond 18 months when stored at -80°C in single-use aliquots, though manufacturer recommendations typically specify -20°C as sufficient.
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