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DLAT Antibody – Rabbit Polyclonal

For Research Use Only. Not for use in diagnostic procedures. Catalog #: MBS1755362

Dihydrolipoyl transacetylase (or dihydrolipoamide acetyltransferase) is an enzyme component of the multienzyme pyruvate dehydrogenase complex. This gene encodes component E2 of the multi-enzyme pyruvate dehydrogenase complex (PDC). PDC resides in the inner… More details

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Product Overview

Online Datasheet in PDF
Product Name
DLAT, Polyclonal Antibody
Full Product Name
Anti-DLAT Antibody Picoband
Product Gene Name
Clonality
Polyclonal
Isotype
Rabbit IgG
Host
Rabbit
Species Reactivity
Human, Mouse, Rat
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Purity/Purification
Immunogen affinity purified.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (lot specific)
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.
Molecular Weight
68,997 Da
OMIM
245348
3D Structure
ModBase 3D Structure for P10515
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human DLAT recombinant protein (Position: P69-P642).
ISO Certification
Manufactured in an ISO 13485:2003 and EN ISO 13485:2012 Certified Laboratory.
Applications Tested/Suitable for anti-DLAT antibody
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes for anti-DLAT antibody
WB: 0.25-0.5 ug/ml, Human, Mouse, Rat
IHC-P: 2-5 ug/ml, Human, Mouse, Rat
ICC/IF: 5 ug/ml, Human
FC/FACS: 1-3 ug/1x10^6 cells, Human
Direct ELISA: 0.1-0.5 ug/ml, Human
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Rabbit IgG (#MBS176460) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (#MBS176453) for IHC(P) and ICC.

Western Blot (WB) of anti-DLAT antibody

Figure 1. Western blot analysis of DLAT using anti-DLAT antibody (MBS1755362).

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 Hela whole cell lysates,
Lane 2: human HEPG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human T-47D whole cell 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-DLAT antigen affinity purified polyclonal antibody (#MBS1755362) at 0.5 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for DLAT at approximately 69 kDa. The expected band size for DLAT is at 69 kDa.
Rabbit Anti-DLAT Polyclonal Antibody – Cat# MBS1755362

Western Blot (WB) of anti-DLAT antibody

Figure 2. Western blot analysis of DLAT using anti-DLAT antibody (MBS1755362).

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: rat brain tissue lysates,
Lane 2: rat kidney tissue lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: mouse kidney tissue lysates,
Lane 5: mouse testis tissue lysates,
Lane 6: mouse NIH/3T3 whole cell 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-DLAT antigen affinity purified polyclonal antibody (#MBS1755362) at 0.5 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for DLAT at approximately 69 kDa. The expected band size for DLAT is at 69 kDa.
Rabbit Anti-DLAT Polyclonal Antibody, Western Blot (WB) – Cat# MBS1755362

Immunohistochemistry (IHC) of anti-DLAT antibody

Figure 3. IHC analysis of DLAT using anti-DLAT antibody (MBS1755362).

DLAT was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Rabbit Anti-DLAT Polyclonal Antibody, Immunohistochemistry (IHC) – Cat# MBS1755362

Immunohistochemistry (IHC) of anti-DLAT antibody

Figure 4. IHC analysis of DLAT using anti-DLAT antibody (MBS1755362).

DLAT was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Rabbit Anti-DLAT Polyclonal Antibody, Immunohistochemistry (IHC) – Cat# MBS1755362

Immunohistochemistry (IHC) of anti-DLAT antibody

Figure 5. IHC analysis of DLAT using anti-DLAT antibody (MBS1755362).

DLAT was detected in a paraffin-embedded section of human renal clear cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Rabbit Anti-DLAT Polyclonal Antibody, Immunohistochemistry (IHC) – Cat# MBS1755362

Immunohistochemistry (IHC) of anti-DLAT antibody

Figure 6. IHC analysis of DLAT using anti-DLAT antibody (MBS1755362).

DLAT was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Rabbit Anti-DLAT Polyclonal Antibody, Immunohistochemistry (IHC) – Cat# MBS1755362

Immunohistochemistry (IHC) of anti-DLAT antibody

Figure 7. IHC analysis of DLAT using anti-DLAT antibody (MBS1755362).

DLAT was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Rabbit Anti-DLAT Polyclonal Antibody, Immunohistochemistry (IHC) – Cat# MBS1755362

Immunofluorescence (IF) of anti-DLAT antibody

Figure 8. IF analysis of DLAT using anti-DLAT antibody (MBS17553627).

DLAT was detected in an immunocytochemical section of Caco-2 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 ug/mL rabbit anti-DLAT Antibody (MBS1755362) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG (MBS177405) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Rabbit Anti-DLAT Polyclonal Antibody, Immunofluorescence (IF) – Cat# MBS1755362

Flow Cytometry (FC/FACS) of anti-DLAT antibody

Figure 9. Flow Cytometry analysis of A431 cells using anti-DLAT antibody (MBS1755362).

Overlay histogram showing A431 cells stained with MBS1755362 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DLAT Antibody (MBS1755362, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG (MBS177405, 5-10 ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Rabbit Anti-DLAT Polyclonal Antibody, Flow Cytometry (FC/FACS) – Cat# MBS1755362

Other Notes

Small volumes of anti-DLAT antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

Searchable Terms for DLAT purchase

MBS1755362 is an antibody targeting DLAT. Also known as Anti-DLAT Antibody Picoband. Reactive in Human, Mouse, Rat. Host: Rabbit.

Additional Details

NCBI/Uniprot data below describe general gene information for DLAT. It may not necessarily be applicable to this product.

NCBI GI #
NCBI GeneID
NCBI Accession #
UniProt Primary Accession #
UniProt Secondary Accession #
UniProt Related Accession #
GenBank Nucleotide #
NCBI Official Full Name
dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial
NCBI Official Synonym Full Names
dihydrolipoamide S-acetyltransferase
NCBI Official Symbol
DLAT
NCBI Official Synonym Symbols
DLTA; PDCE2; PDC-E2
UniProt Protein Name
Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial
UniProt Synonym Protein Names
70 kDa mitochondrial autoantigen of primary biliary cirrhosis; PBC; Dihydrolipoamide acetyltransferase component of pyruvate dehydrogenase complex; M2 antigen complex 70 kDa subunit; Pyruvate dehydrogenase complex component E2; PDC-E2; PDCE2
UniProt Gene Name
DLAT
UniProt Synonym Gene Names
DLTA; PBC; PDC-E2; PDCE2
UniProt Entry Name
ODP2_HUMAN
NCBI Protein Information
dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial; 70 kDa mitochondrial autoantigen of primary biliary cirrhosis; E2 component of pyruvate dehydrogenase complex; M2 antigen complex 70 kDa subunit; PBC

NCBI Summary for DLAT

This gene encodes component E2 of the multi-enzyme pyruvate dehydrogenase complex (PDC). PDC resides in the inner mitochondrial membrane and catalyzes the conversion of pyruvate to acetyl coenzyme A. The protein product of this gene, dihydrolipoamide acetyltransferase, accepts acetyl groups formed by the oxidative decarboxylation of pyruvate and transfers them to coenzyme A. Dihydrolipoamide acetyltransferase is the antigen for antimitochondrial antibodies. These autoantibodies are present in nearly 95% of patients with the autoimmune liver disease primary biliary cirrhosis (PBC). In PBC, activated T lymphocytes attack and destroy epithelial cells in the bile duct where this protein is abnormally distributed and overexpressed. PBC enventually leads to cirrhosis and liver failure. Mutations in this gene are also a cause of pyruvate dehydrogenase E2 deficiency which causes primary lactic acidosis in infancy and early childhood.[provided by RefSeq, Oct 2009]
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UniProt Comments for DLAT

DLAT: the E2 component of the pyruvate dehydrogenase complex (PDHC) that catalyzes the overall conversion of pyruvate to acetyl-CoA and CO2. Contains two lipoyl-binding domains. The PDHC plays a major role in controlling the balance between lipid and glucose oxidation depending on substrate availability. The activity of the PDHC is tightly regulated by phosphorylation of the E1 components by the PDHKs. The pyruvate dehydrogenase (PDH) holoenzyme is a multi-enzyme complex (PDHC) that contains 20-30 copies of pyruvate decarboxylase tetramers (2 alpha:2 beta)(E1), 60 copies of dihydrolipoamide acetyltransferase (E2), six homo-dimers of dihydrolipoamide dehydrogenase (E3), plus E3 binding proteins. PDHKs are recruited to the PDHC by binding to a lipoyl group covalently attached to K174 of the inner lipoyl domain of the E2 component.

Protein type: Carbohydrate Metabolism - pyruvate; Carbohydrate Metabolism - glycolysis and gluconeogenesis; EC 2.3.1.12; Transferase; Carbohydrate Metabolism - citrate (TCA) cycle; Mitochondrial

Chromosomal Location of Human Ortholog: 11q23.1

Cellular Component: mitochondrion; mitochondrial matrix; mitochondrial pyruvate dehydrogenase complex

Molecular Function: dihydrolipoyllysine-residue acetyltransferase activity; protein binding

Biological Process: cellular metabolic process; tricarboxylic acid cycle; glucose metabolic process; regulation of acetyl-CoA biosynthetic process from pyruvate; pyruvate metabolic process

Disease: Pyruvate Dehydrogenase E2 Deficiency
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Precautions

All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines.

Disclaimer

While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.

It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.

Product Note

Select online data sheet information is drawn from bioinformatics databases, occasionally resulting in ambiguous or non-relevant product information. It is the responsibility of the customer to review, verify, and evaluate the information to make sure it matches their requirements before purchasing the kit. Our ELISA Kit assays are dynamic research tools and sometimes they may be updated and improved. If the format of this assay is important to you then please request the current manual or contact our technical support team with a presales inquiry before placing an order. We will confirm the current details of the assay. We cannot guarantee the sample manual posted online is the most current manual, it is intended to serve as an example only. Please refer to the instructions For Use provided with the assay kit for precise details.

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