Anti-NTHL1 Antibody Picoband™

Product Name

Anti-NTHL1 Antibody Picoband™

View all NTH1 Antibodies

SKU/Catalog Number

A03207-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-NTHL1 Antibody Picoband™ catalog # A03207-2. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human.

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-NTHL1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03207-2)

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 NTHL1 recombinant protein (Position: H52-L312).

*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

A03207-2 is reactive to NTHL1 in Human

Applications

A03207-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

34 kDa

Calculated molecular weight

Background of NTH1

Endonuclease III-like protein 1 is an enzyme that in humans is encoded by the NTHL1 gene. The protein encoded by this gene is a DNA N-glycosylase of the endonuclease III family. Like a similar protein in E. coli, the encoded protein has DNA glycosylase activity on DNA substrates containing oxidized pyrimidine residues and has apurinic/apyrimidinic lyase activity.

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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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, 0.1-0.25 µg/ml, Human
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

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Figure 1. Western blot analysis of NTHL1 using anti-NTHL1 antibody (A03207-2).
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 293T whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human A549 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-NTHL1 antigen affinity purified polyclonal antibody (Catalog # A03207-2) at 0.25 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 NTHL1 at approximately 34 kDa. The expected band size for NTHL1 is at 34 kDa.

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Figure 2. IF analysis of NTHL1 using anti-NTHL1 antibody (A03207-2) and anti-Beta Tubulin antibody (M01857-3).
NTHL1 was detected in immunocytochemical section of HELA cell. 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-NTHL1 Antibody (A03207-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of MCF-7 cells using anti-NTHL1 antibody (A03207-2).
Overlay histogram showing MCF-7 cells stained with A03207-2 (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-NTHL1 Antibody (A03207-2, 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 NTHL1 (Source: Uniprot.org, NCBI)

Alternative Names

EC 4.2.99.18; endonuclease III-like protein 1; nth endonuclease III-like 1 (E. coli); NTH1; NTHL1; OCTS3; OCTS3NTH1nth (E.coli endonuclease III)-like 1 NTHL1 FAP3, NTH1, OCTS3, hNTH1 nth like DNA glycosylase 1 endonuclease III-like protein 1|DNA glycoslyase/AP lyase|DNA glycosylase/AP lyase|bifunctional DNA N-glycoslyase/DNA-(apurinic or apyrimidinic site) lyase|bifunctional DNA N-glycosylase/DNA-(apurinic or apyrimidinic site) lyase|nth endonuclease III-like 1

For more info on NTHL1, check out the NTHL1 Infographic

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In this infographic, you will see the following information for NTHL1: 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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