Anti-NUDT5 Antibody Picoband™

Product Name

Anti-NUDT5 Antibody Picoband™

View all ADP-Sugar Pyrophosphatase/NUDT5 Antibodies

SKU/Catalog Number

A07530

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-NUDT5 Antibody Picoband™ catalog # A07530. Tested in ELISA, IHC, WB, Flow Cytometry applications. This antibody reacts with Human, 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-NUDT5 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A07530)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human NUDT5 recombinant protein (Position: M1-F219).

*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

A07530 is reactive to NUDT5 in Human, Rat

Applications

A07530 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

35 kDa

Calculated molecular weight

24.328kDa

Background of ADP-Sugar Pyrophosphatase/NUDT5

ADP-sugar pyrophosphatase is an enzyme that in humans is encoded by the NUDT5 gene. This gene belongs to the Nudix (nucleoside diphosphate linked moiety X) hydrolase superfamily. The encoded enzyme catalyzes the hydrolysis of modified nucleoside diphosphates, including ADP-ribose (ADPR) and 8-oxoGua-containing 8-oxo-dADP and 8-oxo-dGDP. Protein-bound ADP ribose can be hazardous to the cell because it can modify some amino acid residues, resulting in the inhibition of ATP-activated potassium channels. 8-oxoGua is an oxidized form of guanine that can potentially alter genetic information by pairing with adenine and cytosine in RNA. Presence of 8-oxoGua in RNA results in formation of abnormal proteins due to translational errors.

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, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NUDT5 using anti-NUDT5 antibody (A07530).
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 293T whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human Jurkat whole cell lysates,
Lane 5: rat RH35 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-NUDT5 antigen affinity purified polyclonal antibody (Catalog # A07530) 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 NUDT5 at approximately 35 kDa. The expected band size for NUDT5 is at 35 kDa.

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Figure 2. IHC analysis of NUDT5 using anti-NUDT5 antibody (A07530).
NUDT5 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 μg/ml rabbit anti-NUDT5 Antibody (A07530) 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.

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Figure 3. IHC analysis of NUDT5 using anti-NUDT5 antibody (A07530).
NUDT5 was detected in a paraffin-embedded section of human lung 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 μg/ml rabbit anti-NUDT5 Antibody (A07530) 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.

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Figure 4. Flow Cytometry analysis of 293T cells using anti-NUDT5 antibody (A07530).
Overlay histogram showing 293T cells stained with A07530 (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-NUDT5 Antibody (A07530, 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.

Gene/Protein Information For NUDT5 (Source: Uniprot.org, NCBI)

Alternative Names

EC 3.6.1.-; EC 3.6.1.13; hYSAH1; nucleoside diphosphate linked moiety X-type motif 5; Nucleoside diphosphate-linked moiety X motif 5; nudix (nucleoside diphosphate linked moiety X)-type motif 5; Nudix motif 5; NUDT5; YSA1; YSA1H; YSA1HADP-sugar pyrophosphatase NUDT5 YSA1, YSA1H, YSAH1, hNUDT5 nudix hydrolase 5 ADP-sugar pyrophosphatase|8-oxo-dGDP phosphatase|nuclear ATP-synthesis protein NUDIX5|nudix (nucleoside diphosphate linked moiety X)-type motif 5

For more info on NUDT5, check out the NUDT5 Infographic

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In this infographic, you will see the following information for NUDT5: 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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