Anti-DNA Ligase III/LIG3 Antibody Picoband™

Product Name

Anti-DNA Ligase III/LIG3 Antibody Picoband™

View all DNA Ligase III Antibodies

SKU/Catalog Number

A02741-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-DNA Ligase III/LIG3 Antibody Picoband™ catalog # A02741-2. Tested in ELISA, Flow Cytometry, 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-DNA Ligase III/LIG3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02741-2)

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 DNA Ligase III/LIG3 recombinant protein (Position: D297-D963).

*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

A02741-2 is reactive to LIG3 in Human, Mouse, Rat

Applications

A02741-2 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

113 kDa

Calculated molecular weight

112.907kDa

Background of DNA Ligase III

DNA ligase 3 is an enzyme that, in humans, is encoded by the LIG3 gene. This gene is a member of the DNA ligase family. Each member of this family encodes a protein that catalyzes the joining of DNA ends but they each have a distinct role in DNA metabolism. The protein encoded by this gene is involved in excision repair and is located in both the mitochondria and nucleus, with translation initiation from the upstream start codon allowing for transport to the mitochondria and translation initiation from a downstream start codon allowing for transport to the nucleus. Additionally, alternate transcriptional splice variants, encoding different isoforms, have been characterized.

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
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of DNA Ligase III/LIG3 using anti-DNA Ligase III/LIG3 antibody (A02741-2).
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 HepG2 whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse RAW264.7 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-DNA Ligase III/LIG3 antigen affinity purified polyclonal antibody (Catalog # A02741-2) 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 DNA Ligase III/LIG3 at approximately 113 kDa. The expected band size for DNA Ligase III/LIG3 is at 113 kDa.

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Figure 2. Flow Cytometry analysis of K562 cells using anti-DNA Ligase III/LIG3 antibody (A02741-2).
Overlay histogram showing K562 cells stained with A02741-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNA Ligase III/LIG3 Antibody (A02741-2, 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 LIG3 (Source: Uniprot.org, NCBI)

Alternative Names

DNA ligase 3; DNA ligase III; EC 6.5.1.1; LIG2; ligase II, DNA, ATP-dependent; ligase III, DNA, ATP-dependent; Polydeoxyribonucleotide synthase [ATP] 3 LIG3 LIG2alpha, LIG3 DNA ligase 3 DNA ligase 3|ligase II, DNA, ATP-dependent|ligase III, DNA, ATP-dependent|polydeoxyribonucleotide synthase [ATP] 3

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