KIF17 Antibody

What is KIF17 and what cellular functions does it serve?

Beyond neuronal functions, KIF17 has been implicated in spermiogenesis. Studies provide evidence that KIF17 and its associated protein ACT may be involved in sperm nuclear reshaping and tail formation during spermatid remodeling . Additionally, KIF17 has been shown to directly regulate microtubule dynamics and stability, with its motor and tail domains having distinct effects on microtubule polymerization .

What types of KIF17 antibodies are available for research applications?

Several types of KIF17 antibodies are available for research, each with specific characteristics and applications:

• Monoclonal antibodies:

  • KIF17 Antibody (D-8): A mouse monoclonal IgG1 kappa light chain antibody that detects KIF17 protein from mouse, rat, and human origins

  • KIF17 Antibody (B-2): A mouse monoclonal IgG1 antibody specifically designed for human samples

• Polyclonal antibodies:

  • Rabbit polyclonal antibodies like the one from antibodies-online (catalog #ABIN7239149) that recognize human KIF17

  • Recombinant rabbit antibodies such as Proteintech's 85046-1-RR that show reactivity with human, mouse, and rat samples

• Available formats include:

  • Non-conjugated antibodies for standard applications

  • Conjugated forms including agarose, horseradish peroxidase (HRP), phycoerythrin (PE), fluorescein isothiocyanate (FITC), and multiple Alexa Fluor® conjugates

The selection of antibody type depends on the specific research application, target species, and experimental design requirements.

What applications can KIF17 antibodies be used for in research?

KIF17 antibodies support multiple experimental applications across various research contexts:

These applications enable researchers to investigate KIF17 expression, regulation, protein interactions, and cellular localization under various experimental conditions, advancing our understanding of its role in neuronal function and other biological processes.

How should I design experiments to study KIF17 transport in neurons?

Designing experiments to study KIF17-mediated transport requires careful consideration of multiple technical approaches:

• Visualization strategies:

  • Fluorescent protein tagging: Expression of YFP-KIF17 fusion proteins allows tracking of KIF17 in living neurons. Studies have shown that YFP-KIF17 expression patterns and localization are similar to endogenous KIF17, with overexpression increasing KIF17 levels by approximately 1.3-fold .

  • Immunofluorescence: Using specific antibodies against KIF17 and its cargo (e.g., NR2B) to visualize their distribution and co-localization.

• Quantitative distribution analysis:

  • Compartmental quantification: Research has shown that YFP-KIF17 distributes predominantly to dendrites (59%) compared to axons (3%), with 38% remaining in the cell body .

  • Three-dimensional reconstruction: This technique reveals precise spatial relationships between KIF17 and synaptic markers like PSD95 .

• Functional perturbation approaches:

  • Antisense oligonucleotides: Treatment with antisense oligonucleotides against KIF17 (sequence: 5′-CAGAGGCTCACCACCGAA-3′) can knockdown KIF17 expression and disrupt NR2B transport .

  • Dominant-negative constructs: Expression of motor domain-deleted KIF17 (e.g., YFP-610) can block KIF17 function without altering endogenous protein levels .

• Cargo identification and validation:

  • Co-immunoprecipitation: Identifying proteins that associate with KIF17 during transport.

  • Co-localization studies: Determining which cargo proteins travel with KIF17-positive vesicles.

These approaches provide comprehensive analysis of KIF17 movement, localization, and function in neuronal transport processes.

What are the recommended dilutions and controls for KIF17 antibody applications?

Optimal dilutions and appropriate controls are critical for successful KIF17 antibody experiments:

Recommended dilutions for Proteintech's KIF17 antibody (85046-1-RR):

Essential controls for KIF17 antibody experiments:

• Positive controls:

  • Tissue samples: Mouse and rat brain tissues are validated positive controls for WB and IHC

  • Transfected cells: Neurons expressing YFP-KIF17 can serve as positive controls for antibody specificity

• Negative controls:

  • KIF17 knockdown: Cells treated with antisense oligonucleotides against KIF17 should show reduced signal

  • Peptide competition: Antibody pre-incubated with immunizing peptide should show reduced binding

  • Non-expressing tissues: Tissues known not to express KIF17 should not show specific signal

• Specificity controls:

  • Evaluate cross-reactivity with KIF17b: Some antibodies against KIF17 also recognize KIF17b

  • Check for other kinesin family members: Verify no cross-reactivity with related proteins like KIF1A

These guidelines help ensure specific and reliable detection of KIF17 in experimental settings.

How can I effectively analyze KIF17's role in NMDA receptor trafficking?

To analyze KIF17's role in NMDA receptor trafficking, researchers can implement several methodological approaches:

• Quantitative assessment of synaptic NMDA receptors:

  • Immunofluorescence analysis of NR2B clusters following KIF17 manipulation

  • Research has shown that neurons treated with antisense oligonucleotides against KIF17 exhibit a 24.8 ± 3.6% decrease in synaptic NR2B clusters

  • Interestingly, the same treatment produces an 18.7 ± 3.3% increase in synaptic NR2A clusters, suggesting compensatory mechanisms

• KIF17 knockdown approaches:

  • Antisense oligonucleotides treatment: Complete inhibition of KIF17 expression reduces NR2B expression by 33.5 ± 2%

  • This treatment also reduces mLin10 expression by 64.3 ± 5.7%, suggesting interconnected regulatory networks

  • Dominant-negative expression: Overexpression of tail domain constructs that lack motor function

• Co-transport visualization:

  • Live imaging of fluorescently tagged KIF17 and NR2B to track co-transport in real time

  • Three-dimensional reconstruction to determine precise spatial relationships between transport vesicles and synaptic structures

• Receptor turnover studies:

  • Analyzing the contribution of KIF17 to the normal turnover of synaptic NMDA receptors

  • Previous studies indicate that NR1 subunits have approximately 22% turnover in 16 hours

• Regulation analysis:

  • Investigating how activity patterns influence KIF17-mediated transport

  • Examining phosphorylation or other post-translational modifications that might regulate KIF17 transport function

These approaches provide comprehensive insights into how KIF17 contributes to NMDA receptor localization and synaptic function.

How can I investigate KIF17's role in regulating microtubule dynamics?

KIF17 has been identified as a direct regulator of microtubule dynamics and stability, with both its motor and tail domains having distinct effects . To investigate this function:

• In vitro microtubule polymerization assays:

  • Using purified KIF17 domains to assess direct effects on microtubule growth

  • The KIF17 motor domain is sufficient to regulate microtubules, though its activity is modulated by EB1 and the KIF17 tail domain

  • Prepare fresh KIF17 protein fragments for each experiment, as they "tended to aggregate and degrade upon freezing and thawing"

• Plus-end tracking studies:

  • KIF17 is targeted to microtubule plus-ends by EB1

  • Visualization of this interaction provides insights into how KIF17 influences microtubule growth dynamics

• Stability assessment:

  • Analyze microtubule acetylation as a marker of stability

  • Use antibodies against acetylated tubulin (such as clone 6-11B-1, Sigma) to quantify stability changes

  • Compare wild-type, knockdown, and domain-specific mutant conditions

• Structure-function analysis:

  • Create domain-specific KIF17 constructs to determine which regions are responsible for microtubule regulatory functions

  • Assess how motor and tail domains cooperate or function independently

• Co-factor identification:

  • Identify proteins like EB1 that modulate KIF17's effects on microtubules

  • Investigate whether these interactions are regulated by cellular signaling pathways

This comprehensive approach allows researchers to distinguish KIF17's transport functions from its direct regulatory effects on the cytoskeleton.

What methods can be used to study KIF17 gene regulation and expression?

Understanding KIF17 gene regulation provides insights into how this protein's expression is controlled in different contexts. Several approaches can be employed:

• Promoter analysis:

  • KIF17 promoter contains binding sites for transcription factors including NRF-1 and NRF-2α

  • Specific promoter regions can be analyzed using primers targeting:

    • Kif17 NRF-1A&B region: −87 to +112 (F: 5′-TGACGTCACGGAGGTTGC-3′, R: 5′-AAGTGTGCGGGCTGGAAC-3′)

    • Kif17 NRF-2α region: −238 to −64 (F: 5′-CTTACCCTGCCTACCTCTGC-3′, R: 5′-CTGGGCAACCTCCGTGAC-3′)

• Electrophoretic mobility shift assays (EMSAs):

  • In vitro binding assays demonstrate that NRF-1 binds specifically to the KIF17 promoter

  • Competition assays with unlabeled probes and supershift assays with anti-NRF-1 antibodies confirm specificity

• Chromatin immunoprecipitation (ChIP):

  • Verifies in vivo interactions between transcription factors and the KIF17 promoter

  • ChIP assays have confirmed that NRF-1 binds to the KIF17 promoter in neurons

  • β-actin exon 5 can serve as a negative control for these experiments

• Expression analysis across development:

  • Western blotting to track KIF17 expression changes during neuronal maturation

  • Quantification methods using ECL detection and digital scanning for signal quantification

These techniques help uncover the transcriptional mechanisms controlling KIF17 expression, which may be particularly important for understanding developmental regulation of neuronal transport.

How can I study the relationship between KIF17 and spermiogenesis?

Recent research has implicated KIF17 in spermiogenesis, with evidence suggesting roles in sperm nuclear reshaping and tail formation . To investigate this function:

• Localization studies:

  • Immunofluorescence (IF) using KIF17 antibodies to track distribution during spermatid development

  • Research has shown that KIF17 signals are "randomly distributed in the perinuclear cytoplasm throughout the process from early spermiogenesis to mature sperm"

  • The localization pattern changes dynamically, with signals that "initially strengthened, gradually weakened, and finally became concentrated in the tail of mature sperm, especially in the middle piece"

• Co-localization with functional partners:

  • Dual labeling with KIF17 and ACT antibodies

  • In mice, "KIF17b and ACT display coupled intracellular localization in male germ cells"

  • During sperm elongation, "ACT migrates to the cytoplasm along with KIF17b and persists in the sperm tail"

• Functional studies:

  • Gene knockout or RNA interference approaches to confirm proposed models

  • Researchers have noted that their model of KIF17 in spermatid remodeling "needs to be confirmed by techniques that include gene knockout and RNA interference"

• Comparative species analysis:

  • Compare KIF17 function across species to identify conserved mechanisms

  • Studies in Pelophylax esculenta provide complementary insights to mouse models

• Antibody specificity verification:

  • Ensure antibodies can distinguish between KIF17 and KIF17b if necessary

  • Check if antibodies recognize species-specific variants of KIF17 in reproductive tissues

These approaches help establish KIF17's contribution to male reproductive biology alongside its better-characterized neuronal functions.

Why might I observe different molecular weights for KIF17 in Western blot experiments?

Researchers frequently encounter variations in KIF17's apparent molecular weight on Western blots. Several factors explain these discrepancies:

Understanding these factors helps researchers correctly interpret Western blot results and avoid misidentification of KIF17 signals.

What should I do if my KIF17 antibody shows unexpected cross-reactivity or background?

When encountering cross-reactivity or background issues with KIF17 antibodies, several troubleshooting approaches can help:

• Verify antibody specificity:

  • Conduct knockdown experiments using antisense oligonucleotides against KIF17

  • Studies have shown complete inhibition of KIF17 expression using antisense oligonucleotides (sequence: 5′-CAGAGGCTCACCACCGAA-3′)

  • The corresponding sense oligonucleotide (5′-TTCGGTGGTGAGCCTCTG-3′) can serve as a control

• Check for cross-reactivity with related proteins:

  • KIF17 antibodies may detect related kinesin family members

  • Verify no signal changes in other kinesins (e.g., KIF5B, KIF1A) when manipulating KIF17 expression

  • Research has shown that KIF17 antisense treatment does not affect KIF5B or KIF1A expression

• Optimize experimental conditions:

  • Adjust antibody concentration based on recommended dilutions:

    • For Western blot: 1:5000-1:50000 (using Proteintech's 85046-1-RR)

    • For IHC: 1:200-1:800

  • Titrate antibody in each testing system to obtain optimal results

  • For IHC, test different antigen retrieval methods (TE buffer pH 9.0 vs. citrate buffer pH 6.0)

• Use appropriate blocking agents:

  • Optimize blocking to reduce non-specific binding

  • Consider longer blocking times or different blocking agents

• Include competing peptides:

  • Pre-incubate antibody with immunizing peptide as a specificity control

  • Some suppliers offer neutralizing peptides specifically for this purpose

These approaches help establish antibody specificity and improve signal-to-noise ratio in KIF17 detection experiments.

How should I interpret changes in NR2A/NR2B ratio following KIF17 manipulation?

Changes in the NR2A/NR2B ratio following KIF17 manipulation reveal important insights about receptor trafficking and compensatory mechanisms:

• Reciprocal regulation of NR2 subunits:

  • KIF17 knockdown using antisense oligonucleotides reduces NR2B expression by 33.5 ± 2%

  • Simultaneously, the same treatment increases NR2A expression by 24.1 ± 5.2%

  • This pattern suggests a compensatory mechanism to maintain total NMDA receptor levels

• Synaptic localization effects:

  • Antisense treatment reduces synaptic NR2B clusters by 24.8 ± 3.6%

  • Concurrently increases synaptic NR2A clusters by 18.7 ± 3.3%

  • NR2C cluster numbers remain unchanged under the same conditions

• Developmental context:

  • The NR2B-to-NR2A subunit transition is a normal developmental process

  • "NR2B subunits are replaced by NR2A subunits during development"

  • KIF17 manipulation may accelerate this natural transition

• Functional implications:

  • NR2A and NR2B confer different properties to NMDA receptors

  • The shift in ratio likely affects synaptic plasticity mechanisms

  • The balance "probably occurs to maintain an appropriate number of functional NMDARs"

• Related protein changes:

  • KIF17 inhibition also reduces mLin10 expression by 64.3 ± 5.7%

  • This suggests a regulatory network involving KIF17, mLin10, and NMDAR subunits

These findings position KIF17 as a central regulator of NMDA receptor composition, with implications for synaptic function and plasticity.

What emerging applications are being developed for KIF17 antibodies in neuroscience?

KIF17 antibodies are increasingly being utilized in cutting-edge neuroscience research, with several promising directions:

• Neurological disorder investigations:

  • KIF17 dysfunction has been implicated in various neurological disorders

  • Antibodies enable researchers to examine KIF17 expression and localization in disease models

  • Disruptions in KIF17 function can lead to impaired receptor trafficking, potentially contributing to synaptic pathologies

• Activity-dependent transport regulation:

  • Examining how neuronal activity patterns modulate KIF17-mediated transport

  • Antibodies allow visualization of KIF17 redistribution following stimulation protocols

  • May reveal mechanisms linking synaptic activity to receptor availability

• Developmental trajectory analysis:

  • Tracking KIF17 expression and cargo selection across neurodevelopmental stages

  • Understanding the role of KIF17 in the developmental switch from NR2B- to NR2A-containing receptors

  • Potentially informative for neurodevelopmental disorders with synaptic dysfunction

• Combined cytoskeletal regulation and transport:

  • Investigating the dual roles of KIF17 in both cargo transport and direct microtubule regulation

  • Antibodies can help distinguish between these functions in complex cellular environments

  • May provide insights into how neurons coordinate cytoskeletal dynamics with cargo delivery

• Circuit-specific analysis:

  • Examining whether KIF17 function differs across specific neural circuits

  • Combining KIF17 antibodies with circuit-tracing approaches

  • Could reveal specialized transport mechanisms in different functional pathways

These emerging applications position KIF17 antibodies as valuable tools for understanding fundamental aspects of neuronal function and disease mechanisms.

How might advanced imaging techniques enhance KIF17 research?

Advanced imaging technologies are revolutionizing KIF17 research by providing unprecedented insights into its dynamics and functions:

• Super-resolution microscopy:

  • Overcoming the diffraction limit to visualize KIF17-containing vesicles at nanoscale resolution

  • Precisely mapping KIF17 localization relative to synaptic structures

  • Building on existing 3D reconstruction approaches that have revealed "YFP-KIF17 vesicles are not colocalized with PSD95 clusters"

• Live-cell imaging with improved temporal resolution:

  • Capturing rapid transport events with high-speed imaging

  • Quantifying parameters like transport velocity, directional changes, and pausing behavior

  • Extending current knowledge about KIF17's "dynamic properties" and "function in the transport of NR2B in living mammalian neurons"

• Correlative light and electron microscopy (CLEM):

  • Combining KIF17 immunofluorescence with ultrastructural analysis

  • Revealing the precise subcellular context of KIF17-mediated transport

  • Potentially identifying novel cargo or associated structures

• Fluorescence recovery after photobleaching (FRAP):

  • Measuring KIF17 mobility and binding dynamics within different cellular compartments

  • Assessing how activity or pharmacological manipulations affect KIF17 mobility

  • Complementing existing approaches to understanding KIF17 dynamics

• Single-molecule tracking:

  • Following individual KIF17 motors to determine step size, processivity, and force generation

  • Revealing heterogeneity in KIF17 behavior that might be masked in population studies

  • Building on established quantification showing "3% of YFP-KIF17 is in the axon, whereas 59% is in the dendrite and 38% remains in the cell body"

These advanced imaging approaches promise to reveal new aspects of KIF17 function at unprecedented resolution, enhancing our understanding of neuronal transport mechanisms.

What potential therapeutic targets might emerge from KIF17 research?

KIF17 research is uncovering several promising avenues for therapeutic development:

• Synaptic plasticity modulation:

  • KIF17 is "crucial for synaptic plasticity and memory formation"

  • Targeting KIF17-mediated transport could potentially enhance cognitive function

  • Particularly relevant for disorders with learning and memory deficits

• NMDA receptor composition regulation:

  • KIF17 inhibition shifts the NR2A/NR2B ratio

  • This balance is critical for excitotoxicity vulnerability and synaptic plasticity

  • Selective modulation might provide neuroprotection while preserving physiological functions

• Developmental timing interventions:

  • KIF17 influences the developmental transition from NR2B to NR2A subunits

  • Therapeutic targeting could potentially correct aberrant developmental trajectories

  • Relevant for neurodevelopmental disorders with altered NMDAR composition

• Novel microtubule-stabilizing approaches:

  • KIF17 "promotes microtubule stabilization in epithelial cells"

  • This function could be harnessed for disorders involving cytoskeletal instability

  • May provide alternatives to traditional microtubule-targeting drugs

• Cargo-specific transport modulation:

  • Beyond NR2B, KIF17 transports other important neuronal proteins

  • Selective enhancement or inhibition of specific cargo transport

  • Could address imbalances in receptor or channel distribution

These potential therapeutic targets highlight the clinical relevance of fundamental KIF17 research and suggest multiple pathways for intervention in neurological and psychiatric disorders.