Anti-DHODH Antibody Picoband™ (monoclonal, 4E3)

Product Name

Anti-DHODH Antibody Picoband™ (monoclonal, 4E3)

View all DHODH Antibodies

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.

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

Applications

M04035 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

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.

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Reconsitution

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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

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). 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 (Red line) was also used as a control.

Gene/Protein Information For DHODH (Source: Uniprot.org, NCBI)

Alternative Names

DHOdehase; DHODH; dihydroorotate dehydrogenase; dihydroorotate dehydrogenase, mitochondrial; Dihydroorotate Oxidase; EC 1.3.3.1; EC 1.3.5.2; human complement of yeast URA1; POADS; URA1 DHODH DHOdehase, POADS, URA1 dihydroorotate dehydrogenase (quinone) dihydroorotate dehydrogenase (quinone), mitochondrial|dihydroorotate oxidase|human complement of yeast URA1

For more info on DHODH, check out the DHODH Infographic

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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].

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