SLC23A3 Antibody

What is SLC23A3 and what is its function in cellular physiology?

SLC23A3 (Solute Carrier Family 23 Member 3) is a transmembrane protein that functions primarily as a sodium-dependent hypoxanthine transporter and may exhibit xanthine-hypoxanthine exchange activity according to functional studies . Gene ontology annotations associate this protein with transmembrane transporter activity . SLC23A3 belongs to the SLC23 family of nucleobase transporters and is sometimes referenced as Na(+)/L-ascorbic acid transporter 3 or Sodium-dependent vitamin C transporter 3 (SVCT3) .

The protein has several aliases in the literature including:

• E2-binding protein 3 (E2BP3)

• HPC E2-binding protein 3

• SVCT3

• Yolk sac permease-like molecule 1 (Yspl1 or YSPL-1)

In humans, SLC23A3 is encoded by a gene with Entrez Gene ID 151295 and corresponds to UniProt accession Q6PIS1 .

SLC23A3 antibodies have been validated for multiple laboratory applications, making them versatile tools for various research contexts . The specific applications vary by antibody product and should be verified before designing experiments.

When selecting an antibody for a specific application, researchers should review validation data and consider factors such as sample preparation requirements and detection sensitivity .

How can I validate the specificity of an SLC23A3 antibody in my experimental system?

Validating antibody specificity is crucial for generating reliable research data. For SLC23A3 antibodies, consider implementing the following validation strategies:

Positive and negative controls:

• Positive control: Use cells/tissues known to express SLC23A3 (based on literature)

• Negative control: Implement one or more of the following:

  • Use knockout or knockdown models (siRNA against SLC23A3)

  • Pre-absorption with immunizing peptide

  • Secondary antibody-only controls

Molecular weight verification: Confirm that the detected band in Western blotting corresponds to the expected molecular weight of SLC23A3 (approximately 64.9 kDa for the full-length protein) .

Multiple antibody approach: Use two different antibodies targeting distinct epitopes of SLC23A3 to confirm consistency in staining patterns or band detection .

Recombinant protein: Test antibody against recombinant SLC23A3 protein as a defined target. Commercial recombinant mouse SLC23A3 protein (AA 1-611) with His-tag is available for such validation purposes .

As observed in protocols for other antibody validations, experimenters should not be blinded to sample identity during antibody validation analysis to facilitate proper comparative assessment .

What are the recommended experimental controls when using SLC23A3 antibodies?

Implementing proper controls is essential for distinguishing true signals from artifacts when working with SLC23A3 antibodies:

Negative controls:

• Isotype control: Use a non-specific antibody of the same isotype (typically IgG) and host species as the SLC23A3 antibody

• Secondary antibody only: Omit primary antibody to identify non-specific binding of secondary antibody

• Blocking peptide: Pre-incubate antibody with excess immunizing peptide to demonstrate specificity

Positive controls:

• Known positive sample: Include tissue/cells with established SLC23A3 expression

• Transfection control: Use cells transfected with SLC23A3 expression vector

• Recombinant protein: Include purified SLC23A3 protein as a Western blot control

Technical controls:

• Loading control: Include housekeeping proteins (e.g., HSP90) for Western blots to normalize expression levels

• Subcellular marker controls: For localization studies, include markers for relevant cellular compartments to determine co-localization or distinct localization patterns

Proper storage conditions (typically -80°C) should be maintained for antibodies to preserve functionality, and repeated freeze-thaw cycles should be avoided as noted in handling guidelines .

How do polyclonal and monoclonal SLC23A3 antibodies compare in terms of specificity and sensitivity?

While both polyclonal and monoclonal antibodies have advantages, their selection should be guided by your specific research requirements:

Polyclonal SLC23A3 antibodies:

• Recognize multiple epitopes on SLC23A3, potentially increasing signal strength

• May provide greater sensitivity for low-abundance SLC23A3 detection

• Available options include rabbit-host polyclonal antibodies with various conjugates (e.g., FITC)

• Generally purified using protein G (>95% purity reported for some commercial products)

• May have higher batch-to-batch variation

Monoclonal SLC23A3 antibodies:

• Recognize a single epitope, potentially offering higher specificity

• Provide consistent results with minimal batch-to-batch variation

• Limited information on monoclonal options was provided in the search results

For applications requiring high specificity (such as distinguishing between closely related family members), monoclonal antibodies may be preferable. For applications where sensitivity is paramount (such as detecting low SLC23A3 expression), polyclonal antibodies might offer advantages .

When using FITC-conjugated antibodies, remember that proper storage in glycerol-containing buffer (e.g., 50% glycerol, 0.01M PBS, pH 7.4) helps maintain conjugate stability .

What is the optimal sample preparation method for detecting SLC23A3 in different applications?

Proper sample preparation is critical for successful detection of SLC23A3 across different experimental techniques:

Western Blotting:

• Cell lysate preparation: Use buffer containing protease inhibitors to prevent degradation

• Protein denaturation: Heat samples at 95°C for 5 minutes in reducing sample buffer

• Loading amount: 20-30 μg of total protein per lane is typically sufficient

• Expected molecular weight: Approximately 64.9 kDa for full-length SLC23A3

Immunohistochemistry:

• Fixation: 4% paraformaldehyde is commonly used

• Antigen retrieval: May be necessary depending on fixation method

• Blocking: Use 5-10% normal serum from the same species as the secondary antibody

• Primary antibody dilution: Typically 1:100 to 1:500 (optimize empirically)

• Detection: Both chromogenic and fluorescent detection systems are compatible

Immunofluorescence:

• Fixation: 4% paraformaldehyde for 10-15 minutes at room temperature

• Permeabilization: 0.1-0.5% Triton X-100 for 5-10 minutes

• Blocking: 5% BSA or normal serum

• Primary antibody incubation: Overnight at 4°C or 1-2 hours at room temperature

• For FITC-conjugated antibodies: Protect from light during all steps

ELISA:

• Sample dilution: Optimize based on expected SLC23A3 concentration

• Standard curve: Use recombinant SLC23A3 protein for quantitative analysis

• Blocking: 1-5% BSA in phosphate-buffered saline

• Detection: HRP-conjugated secondary antibody with appropriate substrate

How can I troubleshoot non-specific binding when using SLC23A3 antibodies?

Non-specific binding is a common challenge when working with antibodies. For SLC23A3 antibodies, consider these troubleshooting approaches:

For Western Blotting:

• Increase blocking time/concentration (5% non-fat milk or BSA)

• Increase washing duration and number of washes

• Optimize primary antibody dilution (try more dilute conditions)

• Reduce secondary antibody concentration

• Add 0.1-0.5% Tween-20 to washing and antibody diluent buffers

For Immunohistochemistry/Immunofluorescence:

• Use more stringent blocking with 5-10% normal serum plus 1% BSA

• Include 0.1-0.3% Triton X-100 in antibody diluent to reduce hydrophobic interactions

• Pre-absorb primary antibody with tissue powder from a negative sample

• Optimize fixation time (over-fixation can increase background)

• For FITC-conjugated antibodies, ensure proper storage in preservative-containing buffer (e.g., 0.03% Proclin 300)

For ELISA:

• Increase blocking concentration or time

• Add 0.05% Tween-20 to washing buffer

• Dilute samples in buffer containing 0.1-1% BSA

• Optimize washing protocol (increase frequency and volume)

If non-specific binding persists, consider validating with alternative SLC23A3 antibodies or implementing additional controls to distinguish specific from non-specific signals .

What considerations should be taken when using SLC23A3 antibodies for co-localization studies?

Co-localization studies require careful experimental design to generate reliable data:

Antibody selection:

• Choose primary antibodies raised in different host species (e.g., rabbit anti-SLC23A3 and mouse anti-organelle marker) to allow simultaneous detection

• If using multiple rabbit antibodies, consider directly conjugated antibodies or sequential staining protocols

• For FITC-conjugated SLC23A3 antibodies, select complementary fluorophores with minimal spectral overlap

Controls:

• Single-antibody controls: Stain samples with each primary antibody separately to assess bleed-through

• Negative controls: Include samples known to lack SLC23A3 expression

• Positive controls: Include markers with known subcellular localization patterns

Technical considerations:

• Fixation optimization: Different fixatives may better preserve certain epitopes or structures

• Sample thickness: Thinner sections (5-10 μm) typically provide better resolution

• Image acquisition: Use sequential scanning to minimize cross-talk between channels

• Analysis: Employ quantitative co-localization analysis (Pearson's correlation, Manders' coefficients)

Validation approaches:

• Confirm co-localization using complementary techniques (e.g., subcellular fractionation)

• Consider super-resolution microscopy for detailed localization studies

• Perform biological replicates to ensure consistency of observations

When interpreting results, remember that SLC23A3 gene ontology annotations suggest transmembrane transporter activity, indicating potential localization to plasma membrane and/or organelle membranes .

What are the emerging research areas involving SLC23A3 antibodies?

As research on SLC23A3 continues to evolve, several promising directions are emerging:

Functional characterization:

• Investigating the role of SLC23A3 in hypoxanthine transport in different tissues

• Exploring potential associations with vitamin C transport pathways

• Examining interactions with other transporter proteins

Disease associations:

• Evaluating SLC23A3 expression changes in various pathological conditions

• Investigating potential roles in metabolic disorders or cancer

Technical advancements:

• Development of more specific monoclonal antibodies

• Creation of phospho-specific antibodies to investigate regulatory mechanisms

• Generation of antibodies against specific SLC23A3 isoforms

Model systems:

• Expanding validation to additional species for comparative biology studies

• Developing conditional knockout models to study tissue-specific functions

As with all antibody-based research, continuing validation of reagent specificity remains crucial, and researchers should maintain awareness of both the capabilities and limitations of current SLC23A3 antibodies .

How can I determine the optimal SLC23A3 antibody dilution for my specific application?

Determining the optimal antibody dilution is essential for balancing signal strength and specificity:

Titration approach:

• Prepare a dilution series (e.g., 1:100, 1:250, 1:500, 1:1000, 1:2000)

• Test each dilution on identical samples

• Select the highest dilution that provides clear, specific signal with minimal background

Application-specific recommendations:

• Western blotting: Starting dilutions of 1:500 to 1:1000 are typical

• IHC/IF: Starting dilutions of 1:100 to 1:500 are common

• ELISA: Starting dilutions of 1:1000 to 1:5000 may be appropriate

Considerations:

• Sample type may affect optimal dilution (fresh vs. fixed tissues)

• Detection system sensitivity influences required antibody concentration

• Antibody affinity varies between products and applications

• Storage conditions can affect antibody activity over time