SLC22A5 Antibody, Biotin conjugated
Description
ELISA Development
The biotin-conjugated antibody is optimized for ELISA, enabling precise quantification of SLC22A5 in biological samples. It binds selectively to the 42–142 amino acid region, ensuring specificity .
Functional Studies
SLC22A5’s role in carnitine transport is central to metabolic disorders like primary systemic carnitine deficiency (CDSP). This antibody aids in studying:
-
Carnitine Uptake: SLC22A5 transports carnitine in a sodium-dependent manner .
-
Drug Transport: OCTN2 also mediates uptake of organic cations (e.g., tetraethylammonium) and chemotherapeutic agents (e.g., oxaliplatin) .
Primary Carnitine Deficiency (PCD)
Mutations in SLC22A5 cause PCD, characterized by hypoglycemia, cardiomyopathy, and skeletal myopathy . Screening studies in Chinese newborns identified SLC22A5 variants (e.g., c.1400C>G) as major contributors to PCD .
Mechanistic Insights
-
Carnitine Homeostasis: SLC22A5 is critical for intestinal absorption and renal reabsorption of carnitine .
-
Cholesterol Effects: Cholesterol enhances OCTN2-mediated carnitine uptake, highlighting lipid-protein interactions .
Comparative Performance
| Antibody | Reactivity | Applications | Conjugate | Key Advantages |
|---|---|---|---|---|
| Biotin-conjugated | Human | ELISA | Biotin | High sensitivity, streptavidin compatibility |
| HRP-conjugated | Human | ELISA | HRP | Direct detection without secondary antibodies |
| Unconjugated | Human, Rat, Mouse | WB, IF | None | Flexibility for custom labeling |
References and Further Reading
-
Antibodies-online. (2019). SLC22A5 Antibody Biotin (ABIN7170161).
-
Cusabio. (2025). SLC22A5 Antibody raised in Rabbit.
-
GeneBio Systems. (2022). SLC22A5 Antibody, Biotin Conjugated.
-
PMC. (2021). Cholesterol stimulates the cellular uptake of L-carnitine by the OCTN2 transporter.
-
Hoelzel Biotech. (n.d.). SLC22A5 Antibody, Biotin Conjugated.
-
PMC. (2024). Screening primary carnitine deficiency in 10 million Chinese newborns.
-
Abbexa. (2024). Organic cation/carnitine transporter 2 (SLC22A5) Antibody (Biotin).
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Inhibition of endogenous OCTN2-mediated colistin transport through co-incubation with L-carnitine effectively protected primary mouse proximal tubular cells from colistin toxicity. PMID: 28986476
- The OCTN2 carnitine transporter is indispensable for maintaining carnitine levels within the body, ensuring adequate supply to the heart and skeletal muscle, which primarily utilize fat as an energy source. Mutations affecting OCTN2 function lead to carnitine deficiency, potentially manifesting early in life with hypoketotic hypoglycemia or later in life with cardiomyopathy and sudden death due to cardiac arrhythmia. PMID: 26828774
- A homozygous stop variant in the SLC22A5 gene was identified in a family exhibiting cardiomyopathy and a history of sudden death. PMID: 28295041
- Elucidation of GM-CSF signaling revealed that the cytokine activates mTOR kinase, resulting in the phosphorylation and activation of STAT3, which, in turn, regulates OCTN2 transcription. PMID: 27733576
- The dissociation of bound substrate from the transporter is the rate-limiting step in establishing maximal rates of OCT2-mediated transport. PMID: 28615288
- It is proposed that ZO-1, in its unphosphorylated state by PKC, maintains Octn2 in an active state. Conversely, disruption of this binding in DeltaPDZ mutant or after ZO-1 phosphorylation leads to a reduction in Octn2 activity. PMID: 28257821
- Genetic analysis confirmed the diagnosis of systemic primary carnitine deficiency (CDSP) in eight patients at the gene level, including six mutations identified within the solute carrier family 22 member 5 (SLC22A5) gene. PMID: 28186590
- Our findings suggest that a common promoter haplotype of OCTN2 regulates its transcriptional rate and influences the clinical course of CD. PMID: 26965072
- The local genotype influences methylation levels at SLC22A5 and ZPBP2 promoters independently of asthma status. Further research is necessary to confirm the relationship between GSDMA-ZPBP2 and SLC22A5 methylation and asthma in females and males separately. PMID: 26671913
- The results of the current study demonstrated that the -207C>G polymorphism of the SLC22A5 gene is not associated with male infertility. PMID: 26370461
- The c.760C>T (p.R254X) mutation of the SLC22A5 gene has been linked to primary carnitine deficiency. PMID: 26252091
- Human OCTN2 expression is directly regulated by PPAR-alpha. PMID: 25299939
- Nine novel SLC22A5 gene mutations were identified and characterized in Chinese patients with Systemic primary carnitine deficiency (CDSP). The R254X mutation was the most prevalent, and it is likely an ethnic founder mutation. PMID: 25132046
- A novel in-frame deletion (p.F23del), and a novel nonsense mutation (p.Q180X) have been implicated in primary carnitine deficiency. PMID: 23379544
- Mutation analysis of the SLC22A5 gene confirms the diagnosis of primary systemic carnitine deficiency. PMID: 22260907
- OCTN2 is involved in L-carnitine transport at the human blood-brain barrier (BBB). PMID: 23877104
- Mutations in SLC22A5 and ETFDH are associated with riboflavin responsive-multiple acyl-CoA dehydrogenase deficiency. PMID: 25119904
- Promoter methylation is responsible for epigenetic down-regulation of OCTN2 in HepG2 and LS174T cells. PMID: 24146874
- The OCTN2 transporter is generally down-regulated in virus and nonvirus-mediated epithelial cancers, likely due to methylation of its promoter region. PMID: 22374795
- In addition to a significant decrease in free carnitine, carnitine ester metabolism is affected in OCTN2 deficiency in a family with a deletion of 844C of the SLC22A5 gene. PMID: 19238580
- SLC22A5 is an estrogen-dependent gene regulated through a newly identified intronic estrogen response element. PMID: 22212555
- Findings indicate that etoposide can inhibit hOCTN2 function, potentially disrupting carnitine homeostasis. PMID: 22389472
- OCTN2 polymorphisms were not significantly associated with either cancer risk or progression. PMID: 21793125
- Fibroblasts from asymptomatic women have, on average, higher levels of residual carnitine transport activity compared to symptomatic patients due to the presence of at least one missense SLC22A5 mutation. PMID: 21922592
- There is no significant correlation of SLC22A5 polymorphisms with Crohn's disease. PMID: 22118696
- Downregulation of carnitine organic cation transporters 2 is associated with ulcerative colitis. PMID: 21910182
- Impaired plasma membrane targeting of the D122Y and K302E-hOCTN2 variants, which occur in Singaporean populations, contributes to decreased carnitine influx. PMID: 21864509
- The overall expression level of OCTN2 messenger RNA at the inflamed mucosa was significantly reduced compared to noninflamed areas, in both Crohn's disease and ulcerative colitis patients. PMID: 21287663
- OCTN1 and OCTN2 both transport oxaliplatin and are functionally expressed by dorsal root ganglion neurons. PMID: 21606177
- Under hypoxic conditions, placental OCTN2 is down-regulated through PPARalpha-mediated pathways. PMID: 21125992
- OCTN2 mutations are associated with primary carnitine deficiency. PMID: 21126579
- The entire coding regions of the OCTN2 gene were sequenced in 143 unrelated subjects suspected of having Systemic primary carnitine deficiency. PMID: 20574985
- Analysis of plasma carnitine ester profiles in Crohn's disease and ulcerative colitis patients with different IGR2230a_1 genotypes. PMID: 19735486
- OCTN2 possesses functional sites for carnitine and Na(+), and the carnitine-binding site is partially involved in the recognition of organic cations. PMID: 12183691
- Novel missense mutations in the OCTN2 gene (1340A >G and 83G>T) were identified in two Saudi patients with systemic carnitine deficiency. PMID: 12408185
- Studies have demonstrated that l-carnitine uptake in differentiated Caco-2 cells is primarily mediated by OCTN2, located on the brush border membrane. PMID: 12684216
- OCTN2 expression is downregulated in elderly individuals and in patients with myelodysplastic syndrome, with a reduction exceeding 85% compared to younger adults. PMID: 12802501
- Multiple domains of the OCTN2 transporter are necessary for carnitine transport. PMID: 14506273
- Tyrosine residues are involved in coupling the sodium electrochemical gradient to transmembrane solute transfer in the sodium-dependent co-transporter OCTN2. PMID: 14665638
- A G-->C transversion in the promoter of SLC22A5 is associated with Crohn disease. PMID: 15107849
- The reported properties of OCTN2 resemble those observed for l-carnitine uptake in placental brush border vesicles, suggesting that OCTN2 may mediate most maternofetal carnitine transport in humans. PMID: 15238359
- A truncating R254X mutation in the OCTN2 gene was found in a Saudi Arabian kindred, suggesting that it may be a recurrent mutation or a very ancient founder mutation. PMID: 15303004
- OCTN2 is localized in the apical membrane of syncytiotrophoblasts, suggesting a major role in the uptake of carnitine during fetal development. PMID: 15486076
- A homozygous deletion of 17081C of the SLC22A5 gene results in a frameshift at R282D and ultimately leads to a premature stop codon (V295X) in the OCTN2 carnitine transporter in children with cardiomyopathy and decreased plasma carnitine. PMID: 15487009
- Carnitine transport by OCTN2 requires functional linkage between transmembrane domains (TMD) 1-7 and TMD11. PMID: 15499185
- Co-transfection of OCTN2 with PDZK1 stimulated the uptake of carnitine, its endogenous substrate, by OCTN2. PMID: 15523054
- Eight new mutations were identified: V153fsX193, W275X, R289X, 1267del+3_+23, M1I, T232M, T468R. PMID: 15714519
- OCTN2 is expressed in the human heart and can be modulated by drug administration. Furthermore, OCTN2 can contribute to the cardiac uptake of cardiovascular drugs. PMID: 16490820
- None of the four haplotypes present in the SLC22A4/SLC22A5 region in 5q31 showed a significant association with rheumatoid arthritis in a Spanish cohort. PMID: 16652416
- Association of type 1 diabetes with a single nucleotide polymorphism mapping to the SLC22A5 gene. PMID: 16796743
Q&A
What are the primary applications of biotin-conjugated SLC22A5 antibodies in experimental workflows?
Biotin-conjugated SLC22A5 antibodies are primarily deployed in ELISA and immunoprecipitation workflows due to their compatibility with streptavidin-HRP or fluorescent streptavidin probes . The biotin tag provides a stable, high-affinity binding platform for signal amplification, critical for low-abundance transporter detection in tissues like kidney or skeletal muscle . Researchers should validate the antibody’s linear epitope (AA 42-142) against their target species, as cross-reactivity is limited to human samples in most commercial clones .
How do researchers validate the specificity of biotinylated SLC22A5 antibodies?
A three-step validation is recommended:
-
Western blotting: Confirm a single band at ~63 kDa in human lysates (kidney or placenta) .
-
Competitive inhibition: Pre-incubate the antibody with recombinant SLC22A5 (42-142AA) to observe signal reduction in ELISA .
-
Knockdown controls: Use siRNA-mediated SLC22A5 suppression in cell lines to verify loss of immunostaining .
Table 1: Validation Parameters for SLC22A5 Antibodies
How can conflicting data arise when comparing SLC22A5 expression across antibody clones?
Discrepancies often stem from epitope masking or isoform recognition. For example:
-
Antibodies targeting AA 42-142 (extracellular) may fail to detect SLC22A5 in fixed cells due to epitope occlusion .
-
Clones raised against AA 1-180 (cytoplasmic) detect intracellular pools but miss surface-localized transporters .
Solution: Combine IF (using AA 1-180 antibodies) with surface biotinylation assays (using AA 42-142 conjugates) to resolve spatial expression patterns .
What experimental parameters optimize SLC22A5 detection in multiplex assays?
Biotin-conjugated antibodies introduce streptavidin cross-talk in multiplex panels. To mitigate:
-
Blocking: Pre-treat samples with free biotin (100 µg/mL, 30 min) to saturate endogenous biotin receptors .
-
Titration: Reduce antibody concentration to 0.5–1 µg/mL to minimize background in streptavidin-rich tissues (e.g., liver) .
-
Sequential staining: Prioritize SLC22A5 detection before other biotinylated probes to avoid channel bleed-through.
Table 2: Optimized Conditions for Multiplex Assays
| Parameter | Recommendation | Rationale |
|---|---|---|
| Antibody Concentration | 0.5–1 µg/mL | Reduces non-specific binding |
| Incubation Time | 2 hrs at 4°C | Enhances epitope accessibility |
| Streptavidin Probe | Alexa Fluor 647-conjugated | Minimizes spectral overlap |
How do structural polymorphisms in SLC22A5 affect antibody performance?
-
Isoform 2 (truncated at AA 402) lacks the AA 42-142 epitope, leading to false negatives in splice variant studies .
-
Post-translational modifications: O-glycosylation at AA 91 can block antibody binding in cancer cell lines .
Workaround: Use deglycosylation enzymes (e.g., PNGase F) during sample preparation or select antibodies targeting C-terminal epitopes (AA 305-354) .
What controls are essential when quantifying SLC22A5 in disease models?
Include:
-
Positive control: HEK293 cells overexpressing SLC22A5 (AA 1-557).
-
Negative control: SLC22A5 KO cell lines (e.g., CRISPR-edited HepG2).
-
Loading control: Co-stain with Na+/K+ ATPase (plasma membrane) or GAPDH (cytosolic) .
How should researchers address low signal-to-noise ratios in immunohistochemistry?
The biotin-streptavidin system amplifies background in lipid-rich tissues. Optimize:
-
Antigen retrieval: Use citrate buffer (pH 6.0, 95°C, 20 min) to expose AA 42-142 .
-
Signal amplification: Pair with tyramide-based systems (e.g., TSATM) at 1:500 dilution .
-
Quenching: Treat sections with 0.3% H2O2/methanol to block endogenous peroxidases .
Can biotin-conjugated SLC22A5 antibodies be used in live-cell imaging?
Yes, but with limitations:
-
Labeling protocol: Incubate live cells with 2 µg/mL antibody for 10 min at 37°C, followed by streptavidin-647 (1:1000) .
-
Pitfalls: Antibody internalization occurs within 20 min, confounding surface vs. intracellular signals .
-
Alternative: Use pH-sensitive fluorescent streptavidin probes (e.g., pHrodo™) to distinguish membrane-bound vs. endocytosed SLC22A5 .
