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Product Info Summary
| SKU: | M04035 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-DHODH Antibody Picoband® (monoclonal, 4E3)
SKU/Catalog Number
M04035
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-DHODH Antibody Picoband® (monoclonal, 4E3) catalog # M04035. Tested in Flow Cytometry, IF, 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
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-DHODH Antibody Picoband® (monoclonal, 4E3) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M04035)
Host
Mouse
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Monoclonal
Clone Number
4E3
Isotype
Mouse IgG2b
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human DHODH, different from the related mouse sequence by four amino acids, and from the related rat sequence by two amino acids.
*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
M04035 is reactive to DHODH in Human, Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
43 kDa
Calculated molecular weight
42.867kDa
Background of DHODH
Dihydroorotate dehydrogenase (DHODH) is an enzyme that in humans is encoded by the DHODH gene on chromosome 16. The protein encoded by this gene catalyzes the fourth enzymatic step, the ubiquinone-mediated oxidation of dihydroorotate to orotate, in de novo pyrimidine biosynthesis. This protein is a mitochondrial protein located on the outer surface of the inner mitochondrial membrane.
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
M04035 is guaranteed for Flow Cytometry, 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
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human A431 whole cell, human HepG2 whole cell, human MCF-7 whole cell, rat testis tissue, rat liver tissue, mouse testis tissue, mouse liver tissue
ICC/IF: MCF-7 cell
FCM: U937 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of DHODH using anti-DHODH antibody (M04035).
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 A431 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat liver tissue lysates,
Lane 7: mouse testis tissue lysates,
Lane 8: mouse liver 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 mouse anti-DHODH antigen affinity purified monoclonal antibody (Catalog # M04035) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for DHODH at approximately 43 kDa. The expected band size for DHODH is at 43 kDa.
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Figure 2. IF analysis of DHODH using anti-DHODH antibody (M04035).
DHODH was detected in an 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 5 μg/mL mouse anti-DHODH Antibody (M04035) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.
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Figure 3. Flow Cytometry analysis of U937 cells using anti-DHODH antibody (M04035).
Overlay histogram showing U937 cells stained with M04035 (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 mouse anti-DHODH Antibody (M04035, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 DHODH (Source: Uniprot.org, NCBI)
Gene Name
DHODH
Full Name
Dihydroorotate dehydrogenase (quinone), mitochondrial
Weight
42.867kDa
Superfamily
dihydroorotate dehydrogenase family
Alternative Names
SUMO-activating enzyme subunit 2; Anthracycline-associated resistance ARX; Ubiquitin-like 1-activating enzyme E1B; Ubiquitin-like modifier-activating enzyme 2; UBA2; SAE2, UBLE1B; HRIHFB2115 DHODH DHOdehase, POADS, URA1 dihydroorotate dehydrogenase (quinone) dihydroorotate dehydrogenase (quinone), mitochondrial|dihydroorotate oxidase|human complement of yeast URA1
*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 DHODH, check out the DHODH 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 DHODH: 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-DHODH Antibody Picoband® (monoclonal, 4E3) (M04035)
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