SLC39A10 Antibody, Biotin conjugated

What is SLC39A10 and why is it significant in biological research?

SLC39A10, also known as ZIP10, is a zinc transporter protein belonging to the SLC39 (ZIP) family that mediates zinc uptake across cellular membranes. It contains multiple transmembrane domains, a relatively long extracellular N-terminal sequence, and a long intracellular loop .

SLC39A10 plays critical roles in:

• Hematopoiesis (blood cell formation)

• Zinc homeostasis in immune cells (B cells, T cells, macrophages)

• Cell survival mechanisms through zinc-dependent pathways

• Inflammatory response modulation

Research significance:

• SLC39A10 is the master regulator of zinc upregulation in hematopoietic stem cells (HSCs)

• SLC39A10-mediated zinc import promotes survival of HSCs during definitive hematopoiesis

• Two single nucleotide polymorphisms (SNPs) in SLC39A10 have been associated with inflammatory bowel disease (IBD)

• SLC39A10 serves as a key zinc importer upon T cell receptor activation

What are the typical applications for biotin-conjugated SLC39A10 antibodies in research?

Biotin-conjugated SLC39A10 antibodies provide versatility in experimental applications due to the strong biotin-avidin interaction. Key applications include:

It's essential to optimize dilutions for each specific application and include appropriate controls to validate specificity .

How do I select the appropriate SLC39A10 antibody for my specific research model?

Selection criteria should be based on:

• Epitope location: For studying membrane dynamics, select antibodies targeting extracellular epitopes (e.g., residues 259-273 of mouse SLC39A10 or 41-317 of human SLC39A10)

• Species reactivity: Verify cross-reactivity with your model organism:

  • Human-specific: ABIN2781666, H00057181-M03

  • Multi-species (human, mouse, rat): A09043, AZT-010

  • Extended reactivity (including additional species): Check predicted reactivity tables in product datasheets

• Validation status: Prioritize antibodies validated in your application of interest:

  • Western blot: Most SLC39A10 antibodies detect bands at ~68-94 kDa

  • Flow cytometry: Extracellular epitope antibodies perform well in live cell assays

  • Immunohistochemistry: Confirmed reactivity in tissue sections with appropriate controls

• Conjugation compatibility: For biotin-conjugated options, ensure the conjugation doesn't interfere with the antibody's binding capacity

What protocols are recommended for detecting SLC39A10 expression in different cell types using biotin-conjugated antibodies?

Protocol variations by cell type:

For hematopoietic stem cells (HSCs):

• Use single-cell suspension from bone marrow or umbilical cord blood

• Block with 2% BSA in PBS for 30 minutes

• Apply biotin-conjugated SLC39A10 antibody (1:100) for 1 hour at 4°C

• Wash 3× with PBS + 0.1% Tween-20

• Detect with streptavidin-conjugated fluorophore

• Co-stain with HSC markers (e.g., CD34, CD38, CD90) for flow cytometry

For T cells and macrophages:

• Isolate cells using appropriate lineage markers

• Stimulate with relevant activators (LPS for macrophages, anti-CD3/CD28 for T cells)

• Fix with 2% paraformaldehyde (10 minutes) if analyzing intracellular epitopes

• Apply biotin-SLC39A10 antibody at 1:200 dilution

• For flow cytometry, use streptavidin-APC for T cells or streptavidin-FITC for monocytic cell lines

For tissue sections:

• Use fresh-frozen or FFPE sections (4-6 μm thickness)

• For FFPE: Perform heat-induced epitope retrieval (pH 6.0 citrate buffer)

• Block endogenous biotin using a commercial biotin blocking kit

• Apply primary biotin-conjugated antibody overnight at 4°C

• Detect with streptavidin-HRP and appropriate chromogen

• Counterstain with hematoxylin for nuclear visualization

How can I optimize Western blot detection of SLC39A10 using biotin-conjugated antibodies?

Recommended Western blot optimization procedure:

• Sample preparation:

  • Include protease inhibitors in lysis buffer

  • SLC39A10 forms oligomers; avoid excessive heating (keep at 70°C for 10 minutes)

  • For membrane preparation, use gentle detergents (0.5% NP-40 or 1% digitonin)

• Gel selection and transfer:

  • Use 8-10% gels for optimal resolution of the 68-94 kDa SLC39A10 protein

  • Transfer at lower voltage (30V) overnight at 4°C for efficient transfer of membrane proteins

• Blocking and antibody incubation:

  • Block with 5% non-fat milk in TBST (avoid BSA which may contain endogenous biotin)

  • If using biotin-conjugated primary antibody, dilute to 1:500-1:2000

  • Include 0.1% SDS in antibody diluent to reduce non-specific binding

  • Incubate with streptavidin-HRP (1:10,000) for detection

• Expected bands and verification:

  • Primary band at 68-94 kDa (observe both glycosylated and non-glycosylated forms)

  • Verify specificity using SLC39A10 knockout or knockdown controls

  • Include blocking peptide controls to confirm specificity

• Troubleshooting:

  • If detecting multiple bands, check for protein degradation or use fresh samples

  • For weak signals, extend exposure time or increase antibody concentration

  • To reduce background, add 0.05% Tween-20 to wash buffers and use longer washing steps

What factors affect the specificity of SLC39A10 detection in zinc transporter studies?

Several critical factors influence specificity when studying SLC39A10 among other zinc transporters:

• Antibody cross-reactivity:

  • SLC39 family has 14 members with structural similarities

  • Validate that your antibody doesn't cross-react with other ZIP family members, especially ZIP11

  • Use antibodies raised against unique regions rather than conserved domains

• Sample zinc status:

  • SLC39A10 expression is modulated by ambient zinc concentrations

  • Low zinc upregulates SLC39A10, while excess zinc downregulates it

  • Standardize zinc conditions across experimental groups or measure intracellular zinc simultaneously

• Cell type-specific expression patterns:

  • SLC39A10 expression varies across tissues (thymus, spleen, lymph nodes, brain, kidney)

  • Different cell types show variable baseline expression

  • Include appropriate positive and negative control cell types

• Experimental conditions affecting detection:

  • Thyroid hormone status affects SLC39A10 expression

  • JAK-STAT signaling pathways regulate SLC39A10

  • Document and control for pathway activators or inhibitors that may alter expression

• Technical considerations:

  • For flow cytometry, use live cells for extracellular epitope detection

  • Pre-adsorb antibodies against tissues from SLC39A10 knockout models if available

  • Include isotype controls matched to your biotin-conjugated antibody

How can I use biotin-conjugated SLC39A10 antibodies to investigate zinc-dependent cell survival mechanisms?

Methodological approach for zinc-dependent survival studies:

• Dual parameter analysis of SLC39A10 expression and zinc content:

  • Use biotin-conjugated SLC39A10 antibody with streptavidin-fluorophore A

  • Simultaneously apply zinc-specific fluorescent probe (FluoZin-3)

  • Perform flow cytometry or confocal microscopy for co-localization

  • Quantify correlation between SLC39A10 expression and intracellular zinc levels

• Cell death pathway discrimination:

  • Treat cells with specific inhibitors while monitoring SLC39A10 and zinc:

    • Z-VAD-FMK (apoptosis inhibitor)

    • 3-AM (autophagy inhibitor)

    • Ferrostatin-1 (ferroptosis inhibitor)

    • Necrostatin-1 or GSK'872 (necroptosis inhibitors)

  • Combine with annexin V/propidium iodide staining

  • Correlate SLC39A10 levels with specific death pathway activation

• Genetic manipulation system:

  • Use CRISPR/Cas9 to generate SLC39A10 knockout models

  • Rescue experiments with wild-type or mutant SLC39A10

  • Apply zinc chelator TPEN or zinc supplementation

  • Monitor survival rates and intracellular zinc fluctuations

• Pathway analysis integration:

  • Investigate p53/p21 pathway activation

  • Assess RIPK3/MLKL expression (necroptosis mediators)

  • Monitor caspase activation in relation to SLC39A10 expression and zinc levels

Research findings from SLC39A10 knockout models:

• SLC39A10-deficient HSCs show impaired colony-forming capacity

• Inhibiting necroptosis via necrostatin-1 partially rescues SLC39A10-deficient HSCs

• RIPK3/MLKL double knockout partially rescues SLC39A10-deficient HSCs

• Zinc supplementation nearly fully rescues SLC39A10-deficient HSCs

What are the best approaches for studying SLC39A10 localization and trafficking using biotin-conjugated antibodies?

Advanced imaging and localization protocols:

• Pulse-chase trafficking studies:

  • Surface-label cells with cleavable biotin reagent

  • Apply streptavidin-fluorophore to detect initial surface expression

  • Allow internalization at 37°C for various time points

  • Strip remaining surface biotin with reducing agent

  • Fix, permeabilize and apply anti-SLC39A10 antibody with different fluorophore

  • Quantify co-localization to track protein movement

• Co-localization with subcellular markers:

  • Use biotin-conjugated SLC39A10 antibody with streptavidin-fluorophore

  • Apply antibodies against compartment markers:

    • Na⁺/K⁺-ATPase (plasma membrane)

    • Calnexin (endoplasmic reticulum)

    • GM130 (Golgi apparatus)

    • LAMP1 (lysosomes)

  • Perform super-resolution microscopy for precise localization

  • Calculate Pearson's correlation coefficient for quantitative assessment

• Live-cell imaging approach:

  • For extracellular epitope antibodies (like AZT-010-F, residues 259-273)

  • Apply Fab fragments of biotin-conjugated antibody

  • Use streptavidin-quantum dots for extended imaging

  • Monitor trafficking in response to zinc depletion/supplementation

  • Correlate with zinc reporter systems (e.g., FRET-based sensors)

• Tissue-specific expression mapping:

  • Apply biotinylated antibodies to tissue sections

  • Use tyramide signal amplification for enhanced sensitivity

  • Perform multi-color imaging with cell type-specific markers

  • Quantify expression levels across different tissue regions

  • Compare normal versus pathological samples

How do I resolve conflicting data between different SLC39A10 antibodies and experimental systems?

Systematic approach to resolving discrepancies:

• Antibody validation hierarchy:

  • Verify recognition of recombinant SLC39A10 protein

  • Test in SLC39A10 knockout/knockdown systems

  • Perform epitope blocking experiments with immunizing peptides

  • Compare multiple antibodies targeting different epitopes

  • Validate results with orthogonal methods (RNA analysis, tagged constructs)

• Species-specific considerations:

  • Human versus mouse SLC39A10 shows 83% sequence homology

  • Specific regions have higher variation that may affect antibody binding

  • Compare results between species-matched systems

  • Document results in a species-specific context

• Expression level disparities across systems:

  • SLC39A10 expression varies by cell type and activation state

  • Use quantitative methods (qPCR) to correlate protein with mRNA levels

  • Consider zinc status of different experimental systems

  • Document growth conditions, particularly zinc concentrations

• Methodological comparison table:

• Resolution framework:

  • Document all variables between conflicting experiments

  • Test hypothesis that specific conditions alter SLC39A10 conformation or processing

  • Develop consensus detection approaches using multiple antibodies

  • Consider post-translational modifications that may affect detection

What are the emerging applications of SLC39A10 antibodies in studying pathological conditions?

Cutting-edge applications in disease research:

• Inflammatory bowel disease (IBD) studies:

  • Two SNPs in SLC39A10 are associated with IBD susceptibility

  • T cell-specific SLC39A10 deletion protects against disease progression

  • Methodological approach:

    • Use biotin-conjugated antibodies for immunophenotyping intestinal T cells

    • Compare SLC39A10 expression between IBD patients and controls

    • Correlate expression with disease severity markers and zinc status

• Experimental autoimmune encephalomyelitis (EAE) research:

  • SLC39A10 loss in T cells protects against EAE progression

  • Study design considerations:

    • Analyze SLC39A10 expression in different T cell subsets (Th1, Th17, Treg)

    • Correlate with zinc homeostasis markers

    • Track T cell infiltration in CNS tissues using multilabel immunofluorescence

• Hematopoietic disorders and anemia:

  • SLC39A10 knockout zebrafish develop severe anemia

  • Research approach:

    • Use biotin-conjugated antibodies in colony-forming assays

    • Analyze bone marrow biopsies from anemic patients

    • Correlate SLC39A10 expression with erythroid developmental markers

    • Investigate therapeutic potential of targeted zinc supplementation

• Cancer research applications:

  • Elevated SLC39A10 expression in certain malignancies

  • ZIP10 is involved in the migration of cancer cells

  • Investigation strategy:

    • Multiplex staining with biotin-SLC39A10 and tumor markers

    • Analyze different cancer stages for expression changes

    • Correlate with JAK-STAT pathway activation in tumors

    • Evaluate zinc metabolism alterations in cancer progression

• Macrophage-mediated inflammatory diseases:

  • SLC39A10 regulates macrophage survival through zinc/p53-dependent axis

  • Lower mortality following LPS stimulation in macrophage-specific SLC39A10 knockout mice

  • Research methodology:

    • Use biotin-conjugated antibodies for macrophage phenotyping

    • Analyze tissue-resident macrophages in inflammatory models

    • Correlate expression with inflammatory cytokine profiles

How can biotin-conjugated SLC39A10 antibodies be integrated into single-cell analysis platforms?

Methodological innovations for single-cell studies:

• Mass cytometry (CyTOF) integration:

  • Conjugate streptavidin with metal isotopes

  • Use biotin-SLC39A10 antibody in multi-parameter panels

  • Include zinc transporters and zinc-dependent proteins

  • Analyze up to 40 parameters simultaneously across immune populations

  • Apply dimensionality reduction tools (t-SNE, UMAP) for cell subset identification

• Single-cell RNA-seq paired with protein detection:

  • Apply CITE-seq or REAP-seq technologies

  • Use biotin-conjugated antibodies with oligonucleotide-tagged streptavidin

  • Correlate SLC39A10 protein levels with transcriptome profiles

  • Identify cellular states associated with SLC39A10 expression changes

• Spatial transcriptomics with protein detection:

  • Apply Visium or Slide-seq platforms with antibody detection

  • Map SLC39A10 expression in tissue context alongside gene expression

  • Resolve cellular neighborhoods and microenvironmental factors

  • Correlate with zinc sensor probes for functional associations

• Super-resolution microscopy applications:

  • Use biotin-conjugated SLC39A10 antibody with small streptavidin-fluorophores

  • Apply STORM or PALM techniques for nanoscale resolution

  • Track SLC39A10 oligomerization and clustering

  • Analyze membrane microdomain associations and protein-protein interactions

What are the best practices for validating SLC39A10 antibody specificity in knockout models?

Comprehensive validation framework:

• Complete knockout validation strategy:

  • Use CRISPR/Cas9 to generate full SLC39A10 knockout cells/animals

  • Apply Western blot across multiple tissues/cell types

  • Test multiple antibodies targeting different epitopes

  • Include wild-type, heterozygous, and homozygous knockout samples

  • Document expected band disappearance at 68-94 kDa

• Conditional knockout approaches:

  • Generate cell type-specific knockouts (e.g., Slc39a10^fl/fl;LysM-Cre for macrophages)

  • Apply biotin-conjugated antibodies in flow cytometry

  • Compare target cells (Cre+) vs. non-target cells (Cre-) as internal controls

  • Verify specificity with decreasing signal intensity in target populations

• Inducible knockdown systems:

  • Implement siRNA or shRNA targeting SLC39A10

  • Establish dose-response and time-course of expression reduction

  • Correlate antibody signal intensity with mRNA reduction

  • Test antibody sensitivity threshold for detecting partial knockdown

• Cross-species validation:

  • Compare antibody reactivity in human, mouse, rat, and zebrafish models

  • Document species-specific variations in recognition patterns

  • Align epitope sequences across species to predict cross-reactivity

  • Validate in species-matched knockout systems

• Reintroduction experiments:

  • Rescue SLC39A10 knockout with wild-type or mutant constructs

  • Test antibody recognition of rescue constructs

  • Distinguish endogenous vs. exogenous protein using epitope tags

  • Verify functional rescue alongside antibody detection

What methodological approaches can resolve the relationship between SLC39A10 expression and zinc transport activity?

Integrated functional assessment strategies:

• Real-time zinc flux measurement:

  • Label cells with FluoZin-3 AM zinc indicator

  • Apply biotin-SLC39A10 antibody with compatible fluorophore

  • Perform live-cell imaging during zinc addition/removal

  • Correlate transporter expression with zinc uptake rates

  • Account for other zinc transporters using specific inhibitors

• Structure-function correlation:

  • Generate point mutations in key residues of SLC39A10

  • Express in knockout background

  • Apply biotin-conjugated antibodies to confirm expression

  • Measure zinc transport activity using radioisotopes or fluorescent indicators

  • Identify functionally critical domains and residues

• Post-translational modification mapping:

  • Use phospho-specific or glycosylation-specific detection methods

  • Correlate modifications with transport activity

  • Apply inhibitors of specific modifications

  • Determine if antibody epitopes are affected by modifications

  • Document modification-dependent activity regulation

• Quantitative correlation table:

• Regulatory network analysis:

  • Document SLC39A10 expression changes under JAK-STAT pathway modulation

  • Correlate with zinc transport activity measurements

  • Apply chromatin immunoprecipitation to identify transcription factor binding

  • Create an integrated model of expression-activity relationships