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Product Info Summary
| SKU: | A01145-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IP, IF, ICC, WB |
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Product info
Product Name
Anti-DBT Antibody Picoband®
SKU/Catalog Number
A01145-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-DBT Antibody Picoband® catalog # A01145-2. Tested in WB, ICC/IF, IP, Flow Cytometry, ELISA applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-DBT Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01145-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human DBT recombinant protein (Position: K46-R462). Human DBT shares 89.7% amino acid (aa) sequence identity with mouse DBT.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
A01145-2 is reactive to DBT in Human, Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Calculated molecular weight
53.487kDa
Background of DBT
The branched-chain alpha-keto acid dehydrogenase complex (BCKD) is an inner-mitochondrial enzyme complex involved in the breakdown of the branched-chain amino acids isoleucine, leucine, and valine. The BCKD complex is thought to be composed of a core of 24 transacylase (E2) subunits, and associated decarboxylase (E1), dehydrogenase (E3), and regulatory subunits. This gene encodes the transacylase (E2) subunit. Mutations in this gene result in maple syrup urine disease, type 2. Alternatively spliced transcript variants have been described, but their biological validity has not been determined.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01145-2 is guaranteed for ELISA, Flow Cytometry, IP, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Immunoprecipitation, 2-4 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of DBT using anti-DBT antibody (A01145-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human SiHa whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human Hacat whole cell lysates,
Lane 4: human Jurkat whole cell lysates,
Lane 5: rat stomach tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse stomach tissue lysates,
Lane 8: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DBT antigen affinity purified polyclonal antibody (A01145-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DBT at approximately 53 kDa. The expected band size for DBT is at 53 kDa.
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Figure 2. IF analysis of DBT using anti-DBT antibody (A01145-2) and anti-Beta Tubulin antibody (M01857-3).
DBT was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-DBT Antibody (A01145-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and DyLight®550 Conjugated Goat Anti-Mouse IgG (BA1133) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 3. Immunoprecipitating DBT in A431 whole cell lysate.
Western blot analysis of DBT using anti-DBT antibody (A01145-2).
Lane 1: A431 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-DBT antibody in A431 whole cell lysate,
Lane 3: anti-DBT antibody (2μg) + A431 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-DBT antigen affinity purified polyclonal antibody (A01145-2) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for DBT at approximately 53 kDa. The expected band size for DBT is at 53 kDa.
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Figure 4. Flow Cytometry analysis of SiHa cells using anti-DBT antibody (A01145-2).
Overlay histogram showing SiHa cells stained with A01145-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DBT Antibody (A01145-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For DBT (Source: Uniprot.org, NCBI)
Gene Name
DBT
Full Name
Lipoamide acyltransferase component of branched-chain alpha-keto acid dehydrogenase complex, mitochondrial
Weight
53.487kDa
Superfamily
2-oxoacid dehydrogenase family
Alternative Names
BCATE2BCKAD E2 subunit; BCKADE2; BCKAD-E2; branched chain acyltransferase, E2 component; Branched-chain alpha-keto acid dehydrogenase complex component E2; Dihydrolipoamide acetyltransferase component of branched-chain alpha-keto aciddehydrogenase complex; dihydrolipoamide branched chain transacylase (E2 component of branched chainketo acid dehydrogenase complex; maple syrup urine disease); dihydrolipoamide branched chain transacylase E2; Dihydrolipoamide branched chain transacylase; dihydrolipoyl transacylase; Dihydrolipoyllysine-residue (2-methylpropanoyl)transferase; E2 component of branche DBT BCATE2, BCKAD-E2, BCKADE2, BCKDH-E2, BCOADC-E2, E2, E2B dihydrolipoamide branched chain transacylase E2 lipoamide acyltransferase component of branched-chain alpha-keto acid dehydrogenase complex, mitochondrial|52 kDa mitochondrial auto of primary biliary cirrhosis|BCKAD E2 subunit|E2 component of branched chain alpha-keto acid dehydrogenase complex|branched chain 2-oxo-acid dehydrogenase complex component E2|branched chain acyltransferase, E2 component|branched-chain alpha-keto acid dehydrogenase complex component E2|dihydrolipoamide acetyltransferase component of branched-chain alpha-keto acid dehydrogenase complex|dihydrolipoyl transacylase|dihydrolipoyllysine-residue (2-methylpropanoyl)transferase|lipoamide acyltransferase component of mitochondrial branched-chain alpha-keto acid dehydrogenase complex|mitochondrial branched chain alpha-keto acid dehydrogenase transacylase subunit (E2b)
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on DBT, check out the DBT Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for DBT: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-DBT Antibody Picoband® (A01145-2)
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