ABCD1 Antibody
Description
Definition
The ABCD1 Antibody is a laboratory reagent designed to detect the ABCD1 protein (ATP-binding cassette subfamily D member 1), a critical transporter for very long-chain fatty acids (VLCFAs) in peroxisomes. This antibody is widely used in molecular biology and clinical research to study ABCD1 function, diagnose associated disorders, and monitor therapeutic interventions.
2.1. Western Blotting
The antibody is commonly employed in Western blotting to quantify ABCD1 protein levels in tissue lysates or cell extracts. For example, studies using rabbit polyclonal antibodies (e.g., CAB12141) have demonstrated high specificity for human ABCD1, facilitating research into peroxisomal fatty acid metabolism .
2.2. Immunohistochemistry (IHC)
ABCD1 antibodies are utilized in IHC to localize the protein within tissues. A 2024 study validated ABCD1 as a diagnostic marker for solid pseudopapillary neoplasms (SPN), achieving 100% specificity and 99.10% sensitivity in differentiating SPN from pancreatic neuroendocrine tumors (NET) .
2.3. Flow Cytometry
Monoclonal antibodies, such as the EPR15929 clone, enable intracellular detection of ABCD1 in flow cytometry, aiding studies of peroxisomal dynamics in immune cells .
3.1. Role in Adrenoleukodystrophy (ALD)
ABCD1 mutations cause X-linked adrenoleukodystrophy, a condition characterized by VLCFA accumulation and neurodegeneration. Antibodies have been used to confirm protein deficiency in patient fibroblasts and monitor therapeutic responses .
3.2. Gene Therapy and Immune Responses
A 2022 case study revealed that gene therapy with ABCD1-transduced stem cells triggered immune responses, with anti-ALDP IgG antibodies detected post-treatment . This highlights the need for pre- and post-therapy antibody screening in ALD patients.
3.3. Diagnostic Markers
ABCD1 expression levels correlate with disease progression in SPN and ALD. A comparative analysis of diagnostic markers is provided below:
Product Specs
Target Background
- A novel heterozygous mutation IVS4+2T>A (c.1393+2T>A) in the ABCD1 gene has been associated with different clinical phenotypes within a family exhibiting adrenoleukodystrophy. PMID: 28601575
- Expression of human ABCD1 in oligodendrocytes has been shown to rescue apoptosis in abcd1 mutant cells. PMID: 28911205
- In X-linked adrenoleukodystrophy, the absence of ABCD1 function results in increased capillary flow heterogeneity in asymptomatic hemizygotes, primarily in white matter regions and developmental stages with a higher probability of progressing to cerebral disease. PMID: 29136088
- As part of its evaluation process, the New York Newborn Screening Program routinely performs Sanger sequencing to identify mutations in the ABCD1 gene. PMID: 27337030
- ABCD1 and ABCD2 play roles in the transport of long and very long chain fatty acids (VLCFA) or their CoA-derivatives into peroxisomes with distinct substrate specificities. Meanwhile, ABCD3 is involved in the transport of branched chain acyl-CoA into peroxisomes. ABCD4 is thought to be involved in the transport of vitamin B12 from lysosomes to the cytosol. PMID: 27766264
- Mutations in SPG11, ATL1, NIPA1, and ABCD1 have been detected in patients with hereditary spastic paraplegia. PMID: 27084228
- Phenotypic variability has been observed in a Tunisian family with X-linked adrenoleukodystrophy caused by the novel p.Gln316Pro mutation. PMID: 26686776
- Childhood cerebral adrenoleukodystrophy (CCALD) is the most prevalent phenotype (64%) among Chinese patients with X-ALD. Eight novel mutations in the ABCD1 gene have been identified and confirmed as disease-causing mutations. PMID: 26454440
- Recent studies demonstrate that a single splicing mutation can affect ABCD1 transcripts and the ALDP protein function. PMID: 25835712
- Both BCAP31 and ABCD1 have been associated with hepatic cholestasis and death before the age of one. Notably, a patient with an isolated deletion at the 3'-end of SLC6A8 exhibited a similarly severe phenotype as seen in BCAP31 deficiency. PMID: 24597975
- Exome sequencing in two brothers with distinct phenotypes, including congenital language disorder, growth retardation, intellectual disability, and urinary and fecal incontinence, revealed missense mutations in ABCD1 and DACH2. PMID: 25234129
- The loss of ABCD1 function leads to a pathogenic accumulation of very long chain fatty acids, ultimately resulting in mitochondrial dysfunction. PMID: 25393703
- The study in question investigates [mention the specific topic or question addressed in the referenced study]. PMID: 25044748
- The same mutation in the ABCD1 gene has been detected in two unrelated patients with X-linked adrenoleukodystrophy. PMID: 25275259
- Four unrelated women have been identified with late-onset progressive spastic paraparesis and heterozygous mutations in the ABCD1 gene. PMID: 24154795
- The pattern of X chromosome inactivation of the ABCD1 gene has been linked to the symptomatic status in female carriers of X-linked adrenoleukodystrophy. PMID: 24480483
- This study conclusively demonstrates that a cryptic splicing-induced aberrant messenger RNA carrying an internal frameshift deletion results from an intronic mutation in the ABCD1 gene. PMID: 23835273
- Array comparative genomic hybridization analysis suggests that the deletion is a genomic rearrangement spanning 90 kb, starting in exon 4 and encompassing ABCD1. PMID: 22994209
- Eight mutations in ABCD1 have been identified, including one novel deletion (c.1477_1488+11del23) and seven known mutations. PMID: 23566833
- In contrast to yeast cells, very long-chain acyl-CoA esters are transported into peroxisomes by ABCD1 independently of additional synthetase activity. PMID: 23671276
- Adrenoleukodystrophy and skewed X chromosome inactivation favoring the mutant ABCD1 allele have been associated with symptom manifestation in heterozygotes from a Chinese pedigree. PMID: 23469258
- Novel mutations in ABCD1 have been identified in unrelated Argentinean X-linked adrenoleukodystrophy patients. PMID: 23300730
- While very long chain fatty acid (VLCFA) is beta-oxidized in the ABCD1-dependent pathway, the ABCD1-independent peroxisomal and mitochondrial beta-oxidation pathways significantly contribute to VLCFA beta-oxidation in astrocytes. PMID: 23123468
- Research indicates that preferential X chromosome inactivation leads to the favored expression of the mutant ABCD1 allele. PMID: 22280810
- A single germline mutation was identified in each index case within the ABCD1 gene. The analysis detected four novel mutations (two missense and two deletion/insertion) and three novel SNPs. Data revealed variable protein expression in different patients. PMID: 21966424
- A standardized conformation-sensitive gel electrophoresis (CSGE) method was employed to detect mutations in the ABCD1 gene in twenty Indian patients with X-ALD. The results were subsequently confirmed by sequencing. An attempt was also made to correlate genotype with phenotype. PMID: 21889498
- Amongst 489 X-linked adrenoleukodystrophy families, 20 cases exhibited de novo mutations in the ABCD1 gene in the index case, indicating that the mutation arose in the maternal germline. PMID: 21700483
- A novel mutation in the ATP-binding domain of the ABCD1 gene has been reported in adrenoleukodystrophy. PMID: 21273699
- HsABCD1 and HsABCD2 demonstrate distinct substrate specificities. PMID: 21145416
- Three female patients heterozygous for ABCD1 gene mutations were reported for the first time in China, and a novel mutation, p.H283R, was identified in this X-linked adrenoleukodystrophy family. PMID: 20376793
- A family carries a novel deletion of one base pair in exon 8 at nucleotide position 2245 (2245delA) in the ABCD1 gene. PMID: 20042197
- Downregulation of ABCD1 may play a role in human renal tumorigenesis. PMID: 19787628
- A contiguous deletion encompassing the X-linked adrenoleukodystrophy gene (ABCD1) and DXS1357E has been linked to a novel neonatal phenotype resembling peroxisomal biogenesis disorders. PMID: 11992258
- Eight novel mutations have been described in this study. PMID: 12175782
- ALDP facilitates the interaction between peroxisomes and mitochondria. When ALDP is deficient in X-ALD, it leads to an increased accumulation of VLCFAs. PMID: 12509471
- Mutations in ABCD1 are distributed heterogeneously across functional domains of ALDP, impacting peroxisomal transport function. PMID: 12530690
- A splice mutation in the 5' end of intron 5 leading to abnormal splicing in exon 5 and exon 6 is implicated as a cause of X-linked recessive adrenoleukodystrophy. PMID: 12579499
- Six distinct missense mutations in ALD were identified in seven Japanese families. PMID: 12624723
- For the first time, mutations in ABCD1 have been identified in Chinese adrenoleukodystrophy patients in mainland China. PMID: 14556192
- There were no hotspot mutations identified in the ABCD1 gene in China. Mutations in the gene were found in over 70% of patients with ALD, but the identified ABCD1 gene mutations did not show a clear correlation between the type of mutation and phenotype. PMID: 14767898
- An ABCA1-independent but cytoskeleton-dependent cholesterol removal pathway might help prevent early atherosclerosis in Tangier disease. PMID: 15001567
- The accumulation of very long-chain fatty acids does not affect mitochondrial function in ABCD1 protein deficiency. PMID: 15772093
- The study explores the PEX19-binding site of the human adrenoleukodystrophy protein. PMID: 15781447
- Adrenomyeloneuropathy should be considered in the differential diagnosis of spastic paraparesis in both men and women. A mutation in the ABCD1 gene has been reported in the French-Canadian population, suggesting the potential for identifying additional cases in the future. PMID: 16018167
- Over half of the mutations (19/34) were located in exon 1 and exon 6, indicating potential hot exons. PMID: 16087056
- Data indicate that fetus 1 possessed the R617G mutation on his ABCD1 gene, making him an adrenoleukodystrophy hemizygote. Fetus 2 did not carry the P534R mutation on his ABCD1 gene and was considered a normal hemizygote. PMID: 16331554
- ABCD1 gene mutations were found in four cases of X-linked adrenoleukodystrophy with elevated VLCFA levels in amniocytes. No mutations were detected in four other cases with normal VLCFA levels in amniocytes. PMID: 17285533
- Mutant ALDPs, harboring mutations in the COOH-terminal half of ALDP, including S606L, R617H, and H667D, underwent degradation by proteasomes after dimerization. The region between transmembrane domain 2 and 3 is essential for targeting ALDP to the peroxisome. PMID: 17542813
- This study examined a patient with adult-onset cerebral form of X-linked adrenoleukodystrophy with dementia of the frontal lobe type, revealing a novel L160P mutation in the ABCD1 gene. PMID: 17662307
- ALDP-encoding mRNA is most abundant in tissues with high energy demands, such as the heart, muscles, liver, and renal and endocrine systems. ALDP is specifically expressed in certain cell types within the brain. PMID: 17761426
Q&A
What is ABCD1 and what cellular functions does it perform?
ABCD1 (ATP-binding cassette sub-family D member 1), also known as ALDP (adrenoleukodystrophy protein), is a critical membrane transporter belonging to the ATP-binding cassette (ABC) transporter superfamily. This protein localizes specifically to the peroxisomal membrane where it serves as a transporter for very long-chain fatty acids (VLCFAs) . The primary function of ABCD1 is to facilitate the transport of VLCFAs into peroxisomes for breakdown through β-oxidation. This 83 kDa protein (calculated molecular weight) consists of 745 amino acids and functions as a half-transporter that dimerizes to form a fully functional complex . When mutations occur in the ABCD1 gene, the resulting dysfunction leads to the accumulation of toxic fatty acids, particularly causing X-linked adrenoleukodystrophy (X-ALD), an inherited neurodegenerative disease affecting the nervous system white matter and adrenal cortex .
What structural features characterize ABCD1 and how do they relate to its function?
Recent cryo-electron microscopy studies have revealed detailed structural insights into ABCD1's functional mechanisms. The protein exists in multiple conformational states during its transport cycle. Six molecular structures of ABCD1 have been identified in four distinct conformational states, including inward-facing and outward-facing configurations . Key structural elements include:
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Transmembrane domains (TMDs) that bind to substrates such as hexacosanoic acid (C26:0)-CoA
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Nucleotide-binding domains (NBDs) that undergo ATP hydrolysis
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A crucial tryptophan residue (W339) on transmembrane helix 5 (TM5) that is essential for substrate binding
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A unique C-terminal coiled-coil domain that negatively modulates ATPase activity
In the substrate transport cycle, ATP molecules pull the two NBDs together, causing the TMDs to open toward the peroxisomal lumen, facilitating substrate release . This structural understanding provides critical insights into how ABCD1 functions and how mutations may disrupt its normal activity.
What types of ABCD1 antibodies are available and how should researchers select the appropriate one?
Researchers have multiple options when selecting ABCD1 antibodies for their investigations:
Selection criteria should include:
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The specific application requirements (Western blot, IHC, IF, etc.)
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Sample type (human vs. mouse tissues/cells)
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Need for monoclonal specificity versus broader polyclonal epitope recognition
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Verified reactivity in tissues or cell lines relevant to the research question
For studies requiring high specificity, the monoclonal antibody may be preferable, while polyclonal antibodies offer versatility across multiple applications and potentially stronger signals due to recognition of multiple epitopes .
What dilution ranges and sample preparation protocols are optimal for different ABCD1 antibody applications?
Optimal dilution ranges vary by application and specific antibody:
Western Blot (WB) Applications:
Immunohistochemistry (IHC) Applications:
Immunofluorescence/Immunocytochemistry (IF/ICC):
Sample Preparation Considerations:
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For IHC applications, antigen retrieval may be performed with TE buffer pH 9.0 or alternatively with citrate buffer pH 6.0
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For cell lysate preparation in Western blotting, HeLa, HepG2, U-87MG, and LO2 cells have been verified as positive controls
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For immunoprecipitation, 0.5-4.0 μg antibody is recommended for 1.0-3.0 mg of total protein lysate
It is always recommended to optimize these conditions for each experimental system as results may be sample-dependent .
How can ABCD1 antibodies be effectively used in diagnostic applications for solid pseudopapillary neoplasms (SPN)?
Recent research has identified ABCD1 as a novel and highly effective diagnostic marker for solid pseudopapillary neoplasms (SPN) of the pancreas. In a comprehensive study employing immunohistochemical staining, ABCD1 demonstrated exceptional efficacy as a diagnostic tool with remarkable specificity and sensitivity metrics (AUC=0.999, sensitivity=99.10%, specificity=100%) .
Methodology for SPN Diagnosis:
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Perform immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections using anti-ABCD1 antibodies
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Evaluate staining intensity using the Histoscore (Hs) method with categorization into four grades:
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Strong positive (+++, 2.5≤Hs≤3)
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Moderate positive (++, 1.5≤Hs≤2.4)
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Weak positive (+, 0.5≤Hs≤1.4)
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Negative (−, 0≤Hs≤0.4)
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Compare results with established patterns: SPNs typically exhibit strong positive (+++) or moderate positive (++) ABCD1 expression, while normal pancreatic tissues show negative or weak positive (+) staining
This approach allows for accurate differentiation of SPN from other pancreatic neoplasms, including neuroendocrine tumors (NET), acinar cell carcinoma (ACC), and pancreatoblastoma (PB), which can be challenging to distinguish based solely on morphological characteristics .
What experimental approaches can be used to investigate ABCD1 function in peroxisomal fatty acid metabolism?
Several methodological approaches can be employed to study ABCD1 function in peroxisomal fatty acid metabolism:
1. Biochemical Analysis of VLCFA Metabolism:
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Measure very long-chain fatty acid (VLCFA) levels in fibroblasts using gas chromatography-mass spectrometry
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Perform D3-C22:0 loading tests to assess D3-C26:0 de novo synthesis and D3-C22:0 β-oxidation
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Evaluate peroxisomal β-oxidation activity as a percentage of control cells (≤34% is highly suspicious for ALD)
2. Immunoblotting for ALDP Protein Detection:
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Western blot analysis using ABCD1 antibodies to detect presence/absence of ALDP protein
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Compare with known controls - absence of ALDP is associated with X-linked adrenoleukodystrophy
3. ATPase Activity Assays:
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Measure ATPase activity of purified ABCD1 (wild-type and mutant forms)
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Typical maximal basal turnover rates range from 29 ± 1 nmol/mg/min for truncated versions to lower rates for full-length ABCD1
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Assess substrate stimulation using C26:0-CoA
4. Structural and Functional Analysis:
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Employ cryo-electron microscopy to determine conformational states during transport cycles
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Analyze ATP binding and hydrolysis using purified protein preparations
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Investigate the effects of mutations on structural integrity and functional capacity
These approaches collectively provide comprehensive insights into ABCD1's role in peroxisomal fatty acid metabolism and can help elucidate the pathophysiological mechanisms underlying X-linked adrenoleukodystrophy.
How can researchers troubleshoot common issues with ABCD1 antibody-based experiments?
When working with ABCD1 antibodies, researchers may encounter several challenges that require specific troubleshooting approaches:
1. Inconsistent Western Blot Results:
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Issue: Variable band intensity or unexpected molecular weight
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Solution: Optimize protein extraction methods specifically for membrane proteins; ABCD1 is a multi-pass membrane protein localized to peroxisomes . Use reducing agents and appropriate detergents to ensure complete solubilization. The expected molecular weight is approximately 75-83 kDa .
2. Weak or Absent Signal in Immunohistochemistry:
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Issue: Poor or no staining in tissue sections
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Solution: Optimize antigen retrieval methods; for ABCD1, TE buffer pH 9.0 is recommended, though citrate buffer pH 6.0 can be used as an alternative . Titrate antibody concentrations carefully, starting with recommended dilutions (e.g., 1:60-1:450 for E-AB-19528) .
3. Specificity Concerns:
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Issue: Potential cross-reactivity with other ABC transporters
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Solution: Include proper negative controls (tissue/cells known to lack ABCD1 expression) and positive controls (HeLa cells, LO2, U-87MG have been verified) . Consider using ABCD1 knockout or knockdown samples as definitive negative controls.
4. Variable Results Across Different Cell Types:
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Issue: Inconsistent expression patterns in different cell lines
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Solution: Cell-specific expression levels may vary naturally. Normalize results to appropriate housekeeping proteins and validate findings using alternative detection methods (qPCR, functional assays).
5. Storage and Handling Issues:
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Issue: Reduced antibody performance over time
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Solution: Store antibodies according to manufacturer recommendations (typically at -20°C with glycerol) . Avoid repeated freeze-thaw cycles, and consider aliquoting antibodies upon receipt.
How can ABCD1 antibodies be used to investigate disease mechanisms in X-linked adrenoleukodystrophy?
ABCD1 antibodies provide valuable tools for investigating disease mechanisms in X-linked adrenoleukodystrophy (X-ALD) through multiple research approaches:
1. Variant Classification and Functional Analysis:
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Use ABCD1 antibodies in conjunction with biochemical assays to classify variants of unknown significance (VUS) in the ABCD1 gene
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Assess ALDP protein expression via Western blotting in patient fibroblasts or peripheral blood mononuclear cells
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Correlate protein expression patterns with VLCFA metabolism metrics to establish pathogenicity of variants
2. Structure-Function Relationship Studies:
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Investigate how specific mutations affect ABCD1 protein localization to peroxisomes using immunofluorescence microscopy
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Examine structural integrity of mutant proteins by comparing immunostaining patterns between wild-type and mutant ABCD1
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Utilize antibodies against different epitopes to identify regions most affected by pathogenic mutations
3. Therapeutic Development and Evaluation:
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Monitor ABCD1 protein expression in response to potential therapeutic interventions
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Assess peroxisomal localization recovery following gene therapy or pharmacological approaches
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Evaluate normalization of peroxisomal function through co-localization studies with other peroxisomal markers
These approaches allow researchers to dissect the molecular pathogenesis of X-ALD and potentially identify novel therapeutic targets or strategies for intervention.
What emerging techniques combine ABCD1 antibody detection with other methodologies for comprehensive protein analysis?
Several cutting-edge methodologies combine ABCD1 antibody detection with other techniques to provide more comprehensive insights:
1. Multi-omics Integration:
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Combine antibody-based protein detection with transcriptomics to correlate ABCD1 protein levels with mRNA expression
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Example: In SPN research, initial gene expression profiling identified ABCD1 as upregulated (Log2FC=1.622 compared to normal tissues and Log2FC=2.057 compared to NET samples), which was subsequently confirmed at the protein level using immunoblotting and immunohistochemistry
2. Advanced Imaging Technologies:
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Super-resolution microscopy combined with ABCD1 immunostaining allows precise localization within peroxisomal membranes
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Techniques such as whole slide image scanning (e.g., NanoZoomer S360, C13220-01) with digital quantification via FIJI software enable objective assessment of staining intensity across multiple regions of interest
3. Structural Biology Applications:
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Antibody-based purification of ABCD1 protein for subsequent structural studies using cryo-electron microscopy
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Detection of specific conformational states using conformation-specific antibodies to complement direct structural studies
4. Functional Assays:
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Combine immunoprecipitation of ABCD1 with ATPase activity measurements to analyze protein function
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Use antibodies to capture ABCD1 protein complexes and identify novel interacting partners through mass spectrometry
These integrated approaches represent the cutting edge of ABCD1 research and will likely contribute significantly to our understanding of its role in health and disease.
