SLC16A11 Antibody, HRP conjugated

What is SLC16A11 and why is it significant for metabolic research?

SLC16A11 (Solute Carrier Family 16 Member 11), also known as Monocarboxylate Transporter 11 (MCT11), is a proton-linked transporter that catalyzes the movement of pyruvate across the plasma membrane . The protein has significant research interest due to its involvement in hepatic lipid metabolism, where overexpression results in increased triacylglycerol (TAG) levels and alterations in other lipid profiles including diacylglycerols, lysophosphatidylcholine, cholesterol ester, and sphingomyelin . Notably, SLC16A11 has been implicated as a causal factor in Type 2 Diabetes (T2D), particularly in Latino populations, making it an important target for metabolic disease research .

What applications are HRP-conjugated SLC16A11 antibodies best suited for?

HRP-conjugated SLC16A11 antibodies are particularly valuable for Western Blotting (WB), ELISA, and specific immunohistochemistry applications . The direct HRP conjugation eliminates the need for secondary antibody incubation, reducing background signal and experimental time. These antibodies provide enhanced sensitivity for detection of low-abundance SLC16A11 in complex tissue samples, especially liver specimens where expression levels can vary significantly based on genetic variants .

How should sample preparation be optimized for SLC16A11 detection in different tissues?

For optimal detection of SLC16A11, tissue-specific extraction protocols should be implemented. For liver samples, where SLC16A11 plays a crucial metabolic role, membrane extraction assays are recommended since approximately 5% of SLC16A11 localizes to the plasma membrane, with the majority found in intracellular membranes . When working with brain tissue, include protease inhibitor cocktails to prevent degradation of the transporter. For all tissues, sample preparation should account for the predicted subcellular localization pattern: predominantly in endoplasmic reticulum with a small fraction at the cell surface .

What dilution ranges are effective for HRP-conjugated SLC16A11 antibodies in different applications?

Based on experimental validation, the following dilution ranges have been determined for optimal signal-to-noise ratio:

These recommendations are based on documented experiments with commercially available antibodies .

How can researchers validate the specificity of SLC16A11 antibodies?

To validate antibody specificity, implement a multi-faceted approach: (1) Compare staining patterns in tissues known to express SLC16A11 (like liver and brain) versus negative control tissues; (2) Perform siRNA knockdown or CRISPR knockout of SLC16A11 in cell lines followed by antibody testing; (3) Use peptide competition assays with the immunizing peptide (AA 48-76 for some commercial antibodies) ; (4) Include samples from individuals with different SLC16A11 genotypes, as expression levels vary in a dose-dependent manner with the T2D risk haplotype .

How can SLC16A11 antibodies be used to investigate the relationship between SLC16A11 and Type 2 Diabetes?

To investigate SLC16A11's role in T2D, researchers can employ HRP-conjugated antibodies in combination with genotyping approaches. Specifically, stratify liver samples from donors based on T2D risk haplotype status (homozygous non-risk, heterozygous, homozygous risk) and quantify SLC16A11 protein levels. This approach complements gene expression studies that have shown 42% lower expression in heterozygotes and 66% lower expression in homozygotes for the T2D risk haplotype . Additionally, immunoprecipitation experiments can be designed to investigate the interaction between SLC16A11 and BSG (basigin), which is disrupted by coding variants in the T2D risk haplotype .

What techniques can be used to study allele-specific effects on SLC16A11 protein expression?

For advanced investigations of allele-specific effects on SLC16A11 protein expression, researchers can implement a comprehensive strategy combining:

• Allelic expression imbalance studies using digital droplet PCR (ddPCR) with probes that distinguish between risk and non-risk alleles

• ChIP-sequencing for histone modifications (H3K27ac, H3K4me1, H3K4me3) in heterozygous samples to analyze the chromatin landscape on each haplotype

• Quantitative immunohistochemistry with HRP-conjugated antibodies to correlate protein levels with genotype

• Pulse-chase experiments to determine if coding variants affect protein stability

Such approaches have revealed that expression from the T2D risk allele is 62% lower than from the non-risk haplotype, indicating a strong cis-effect .

How can researchers investigate the transport function of SLC16A11 using antibody-based approaches?

To study SLC16A11's transport function, combine antibody-based protein detection with functional assays. First, use immunofluorescence or cell surface biotinylation followed by Western blotting with HRP-conjugated antibodies to quantify plasma membrane localization of SLC16A11. Then correlate this with functional transport assays, such as pyruvate uptake measurements using a genetically encoded pyruvate FRET sensor (pyronic) . This multi-modal approach allows researchers to link protein expression and localization with transport activity, particularly important when comparing wild-type SLC16A11 with variants found on the T2D risk haplotype.

What strategies can address non-specific binding when using HRP-conjugated SLC16A11 antibodies?

When encountering non-specific binding with HRP-conjugated SLC16A11 antibodies, implement the following optimization strategies:

• Increase blocking stringency using 5% BSA instead of standard milk blocker

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

• Pre-absorb the antibody with tissue lysate from a species different from your experimental sample

• Optimize antibody concentration through titration experiments, as excessive antibody can increase background

• For IHC applications, perform antigen retrieval optimization, testing both heat-induced and enzymatic methods

These approaches are particularly important when working with tissues that may express proteins with sequence homology to the immunizing peptide (AA 48-76) .

How should researchers address conflicting results between gene expression and protein detection of SLC16A11?

When facing discrepancies between SLC16A11 mRNA and protein levels, consider:

• Post-transcriptional regulation: Examine microRNA binding sites within SLC16A11 transcripts that may affect translation efficiency

• Protein stability differences: Design pulse-chase experiments to determine if variant-specific differences in protein half-life exist

• Subcellular localization changes: Use fractionation protocols to assess if the protein distribution changes between compartments without affecting total expression

• Epitope masking: Test multiple antibodies targeting different regions of SLC16A11, as protein-protein interactions may obscure specific epitopes

• Genetic background effects: Account for the T2D risk haplotype status, which can reduce expression by up to 66% in homozygous carriers

What controls are essential when using SLC16A11 antibodies in studies of variant effects?

When investigating the effects of SLC16A11 variants, the following controls are crucial:

• Genotype-matched samples: Include samples from individuals with known SLC16A11 genotypes spanning non-risk homozygotes, heterozygotes, and risk-haplotype homozygotes

• Expression vectors: Create side-by-side comparisons of cells transfected with wild-type SLC16A11 (SLC16A11-REF) and T2D risk variant-containing SLC16A11 (SLC16A11-T2D)

• Blocking peptide controls: Include controls using the immunizing peptide to validate antibody specificity

• Knockdown/knockout validation: Generate SLC16A11-depleted samples as negative controls

• Cross-reactive protein controls: Test antibody against related SLC16 family members, particularly those with sequence similarity in the antibody epitope region

How can SLC16A11 antibodies contribute to investigations of hepatic lipid metabolism?

HRP-conjugated SLC16A11 antibodies can facilitate innovative research into hepatic lipid metabolism through:

• Co-localization studies with lipid droplet markers in liver sections from individuals with varying metabolic health

• Quantitative analysis of SLC16A11 expression in non-alcoholic fatty liver disease progression

• Proximity ligation assays to identify novel protein interaction partners in hepatocytes

• Correlation of SLC16A11 expression patterns with lipidomic profiles to establish causative relationships

• Chromatin immunoprecipitation studies to identify transcription factors regulating SLC16A11 expression under different metabolic conditions

These approaches can help elucidate the mechanism by which SLC16A11 overexpression increases triacylglycerol levels and affects other lipid species .

What methodological approaches can determine if post-translational modifications affect SLC16A11 function?

To investigate post-translational modifications (PTMs) of SLC16A11:

• Perform immunoprecipitation with SLC16A11 antibodies followed by mass spectrometry to identify PTM sites

• Compare PTM patterns between wild-type and T2D risk variant proteins

• Create site-directed mutants of potential modification sites and assess impacts on transport function

• Use phospho-specific antibodies in combination with general SLC16A11 antibodies to determine regulation by signaling pathways

• Implement 2D gel electrophoresis followed by Western blotting to separate differentially modified forms of SLC16A11

Such studies may reveal how metabolic conditions or genetic variants affect protein function through altered PTM patterns .