Product Info Summary
SKU: | PB9579 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Lipoamide Dehydrogenase/DLD Antibody Picoband®
View all Dihydrolipoamide Dehydrogenase/DLD Antibodies
SKU/Catalog Number
PB9579
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Lipoamide Dehydrogenase/DLD Antibody Picoband® catalog # PB9579. Tested in Flow Cytometry, IF, IHC, ICC, WB 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
Store 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 freeze-thaw cycles.
Cite This Product
Anti-Lipoamide Dehydrogenase/DLD Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9579)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human DLD recombinant protein (Position: K300-F509). Human DLD shares 96.2% and 95.7% amino acid (aa) sequence identity with mouse and rat DLD, respectively.
*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
PB9579 is reactive to DLD in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
54 kDa
Calculated molecular weight
54177 MW
Background of Dihydrolipoamide Dehydrogenase/DLD
DLD, Dihydrolipoamide dehydrogenase, is a component of the pyruvate dehydrogenase complex, the alpha-ketoglutarate dehydrogenase complex, and the branched-chain alpha-keto acid dehydrogenase complex (BCKD). DLD is a flavoprotein enzyme that degrades lipoamide, and produces dihydrolipoamide. The DLD gene contains 14 exons. The gene is localized to 7q31-q32. This gene encodes the L protein of the mitochondrial glycine cleavage system. The L protein, also named dihydrolipoamide dehydrogenase, is also a component of the pyruvate dehydrogenase complex, the alpha-ketoglutarate dehydrogenase complex, and the branched-chain alpha-keto acide dehydrogenase complex.
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
PB9579 is guaranteed for Flow Cytometry, IF, IHC, 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.
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human K562 whole cell, human A431 whole cell, human SiHa whole cell, human U251 whole cell, rat liver tissue, rat heart tissue, mouse liver tissue, mouse heart tissue
IHC: human intestinal cancer tissue
ICC/IF: MCF-7 cell
FCM: A549 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 2. IHC analysis of DLD using anti-DLD antibody (PB9579).
DLD was detected in a paraffin-embedded section of human intestinal 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 1 μg/ml rabbit anti-DLD Antibody (PB9579) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 1. Western blot analysis of DLD using anti-DLD antibody (PB9579).
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 K562 whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse heart 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-DLD antigen affinity purified polyclonal antibody (Catalog # PB9579) 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 DLD at approximately 54 kDa. The expected band size for DLD is at 54 kDa.
Click image to see more details
Figure 3. IF analysis of DLD using anti-DLD antibody (PB9579).
DLD was detected in immunocytochemical section of MCF-7 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 2μg/mL rabbit anti-DLD Antibody (PB9579) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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 4. Flow Cytometry analysis of A549 cells using anti-DLD antibody (PB9579).
Overlay histogram showing A549 cells stained with PB9579 (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-DLD Antibody (PB9579, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 5. Western blot analysis of DLD using anti-DLD antibody (PB9579).
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 K562 whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse heart 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-DLD antigen affinity purified polyclonal antibody (PB9579) 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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for DLD at approximately 54 kDa. The expected band size for DLD is at 54 kDa.
Protein Target Info & Infographic
Gene/Protein Information For DLD (Source: Uniprot.org, NCBI)
Gene Name
DLD
Full Name
Dihydrolipoyl dehydrogenase, mitochondrial
Weight
54177 MW
Superfamily
class-I pyridine nucleotide-disulfide oxidoreductase family
Alternative Names
Dihydrolipoyl dehydrogenase, mitochondrial;1.8.1.4;Dihydrolipoamide dehydrogenase;Glycine cleavage system L protein;DLD;GCSL, LAD, PHE3; DLD DLDDH, E3, GCSL, LAD, OGDC-E3, PHE3, DLD dihydrolipoamide dehydrogenase dihydrolipoyl dehydrogenase, mitochondrial|E3 component of pyruvate dehydrogenase complex, 2-oxo-glutarate complex, branched chain keto acid dehydrogenase complex|diaphorase|epididymis secretory sperm binding protein|glycine cleavage system L protein|glycine cleavage system protein L|lipoamide dehydrogenase|lipoamide reductase|lipoyl dehydrogenase
*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 DLD, check out the DLD 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 DLD: 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-Lipoamide Dehydrogenase/DLD Antibody Picoband® (PB9579)
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Customer Q&As
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1 Customer Q&As for Anti-Lipoamide Dehydrogenase/DLD Antibody Picoband®
Question
We are currently using anti-Lipoamide Dehydrogenase/DLD antibody PB9579 for rat tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on horse tissues as well?
A. Parker
Verified customer
Asked: 2017-08-28
Answer
The anti-Lipoamide Dehydrogenase/DLD antibody (PB9579) has not been tested for cross reactivity specifically with horse tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-08-28