Product Info Summary
SKU: | A05658-1 |
---|---|
Size: | 100 µg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-PDK3 Antibody Picoband™
SKU/Catalog Number
A05658-1
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-PDK3 Antibody Picoband™ catalog # A05658-1. Tested in ELISA, IF, IHC, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
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-PDK3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05658-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human PDK3 recombinant protein (Position: Y84-Q406). Human PDK3 shares 97.2% amino acid (aa) sequence identity with mouse PDK3.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
A05658-1 is reactive to PDK3 in Human, Mouse, Rat
Applications
A05658-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
47 kDa
Calculated molecular weight
52588 MW
Background of PDK3
Pyruvate dehydrogenase lipoamide kinase isozyme 3, mitochondrial is an enzyme that in humans is encoded by the PDK3 gene. The pyruvate dehydrogenase (PDH) complex is a nuclear-encoded mitochondrial multienzyme complex that catalyzes the overall conversion of pyruvate to acetyl-CoA and CO(2). It provides the primary link between glycolysis and the tricarboxylic acid (TCA) cycle, and thus is one of the major enzymes responsible for the regulation of glucose metabolism. The enzymatic activity of PDH is regulated by a phosphorylation/dephosphorylation cycle, and phosphorylation results in inactivation of PDH. The protein encoded by this gene is one of the three pyruvate dehydrogenase kinases that inhibits the PDH complex by phosphorylation of the E1 alpha subunit. This gene is predominantly expressed in the heart and skeletal muscles. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.
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.
Assay dilution & Images
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
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
Immunohistochemistry, 2-5 µg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human
ELISA, 0.1-0.5 µg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of PDK3 using anti-PDK3 antibody (A05658-1).
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 MCF-7 whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human SiHa whole cell lysates,
Lane 5: human Jurkat whole cell lysates,
Lane 6: human U251 whole cell lysates,
Lane 7: human A549 whole cell lysates,
Lane 8: rat testis tissue lysates,
Lane 9: mouse testis 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-PDK3 antigen affinity purified polyclonal antibody (Catalog # A05658-1) 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 PDK3 at approximately 47 kDa. The expected band size for PDK3 is at 47 kDa.
Click image to see more details
Figure 2. IHC analysis of PDK3 using anti-PDK3 antibody (A05658-1).
PDK3 was detected in a paraffin-embedded section of human meningioma 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 μg/ml rabbit anti-PDK3 Antibody (A05658-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of PDK3 using anti-PDK3 antibody (A05658-1).
PDK3 was detected in a paraffin-embedded section of human rectum adenocarcinoma 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 μg/ml rabbit anti-PDK3 Antibody (A05658-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of PDK3 using anti-PDK3 antibody (A05658-1).
PDK3 was detected in a paraffin-embedded section of mouse testis 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 μg/ml rabbit anti-PDK3 Antibody (A05658-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of PDK3 using anti-PDK3 antibody (A05658-1).
PDK3 was detected in a paraffin-embedded section of rat testis 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 μg/ml rabbit anti-PDK3 Antibody (A05658-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 6. IF analysis of PDK3 using anti-PDK3 antibody (A05658-1).
PDK3 was detected in an immunocytochemical section of HELA 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-PDK3 Antibody (A05658-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was 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.
Click image to see more details
Figure 7. Flow Cytometry analysis of U251 cells using anti-PDK3 antibody (A05658-1).
Overlay histogram showing U251 cells stained with A05658-1 (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-PDK3 Antibody (A05658-1, 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 (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For PDK3 (Source: Uniprot.org, NCBI)
Gene Name
PDK3
Full Name
[Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 3, mitochondrial
Weight
52588 MW
Superfamily
PDK/BCKDK protein kinase family
Alternative Names
EC 2.7.11.2; Pyruvate dehydrogenase kinase isoform 3; pyruvate dehydrogenase kinase, isoenzyme 3; pyruvate dehydrogenase kinase, isozyme 3; pyruvate dehydrogenase, lipoamide, kinase isozyme 3, mitochondrial PDK3 CMTX6, GS1-358P8.4 pyruvate dehydrogenase kinase 3 pyruvate dehydrogenase kinase, isozyme 3|[Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 3, mitochondrial|pyruvate dehydrogenase kinase, isoenzyme 3|pyruvate dehydrogenase, lipoamide, kinase isozyme 3, mitochondrial
*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 PDK3, check out the PDK3 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 PDK3: 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-PDK3 Antibody Picoband™ (A05658-1)
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