Anti-DNA Ligase IV/LIG4 Antibody Picoband™

Product Name

Anti-DNA Ligase IV/LIG4 Antibody Picoband™

View all DNA Ligase IV Antibodies

SKU/Catalog Number

A01685-3

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-DNA Ligase IV/LIG4 Antibody Picoband™ catalog # A01685-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB 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-DNA Ligase IV/LIG4 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01685-3)

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 IV/LIG4 recombinant protein (Position: M1-K876).

*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

A01685-3 is reactive to LIG4 in Human

Applications

A01685-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

104 kDa

Calculated molecular weight

103.971kDa

Background of DNA Ligase IV

DNA ligase 4 is an enzyme that in humans is encoded by the LIG4 gene. The protein encoded by this gene is a DNA ligase that joins single-strand breaks in a double-stranded polydeoxynucleotide in an ATP-dependent reaction. This protein is essential for V(D)J recombination and DNA double-strand break (DSB) repair through nonhomologous end joining (NHEJ). This protein forms a complex with the X-ray repair cross complementing protein 4 (XRCC4), and further interacts with the DNA-dependent protein kinase (DNA-PK). Both XRCC4 and DNA-PK are known to be required for NHEJ. The crystal structure of the complex formed by this protein and XRCC4 has been resolved. Defects in this gene are the cause of LIG4 syndrome. Alternatively spliced transcript variants encoding the same protein have been observed.

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
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 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 DNA Ligase IV/LIG4 using anti-DNA Ligase IV/LIG4 antibody (A01685-3).
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 Ramos whole cell lysates,
Lane 2: human HepG2 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 IV/LIG4 antigen affinity purified polyclonal antibody (Catalog # A01685-3) 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 IV/LIG4 at approximately 104 kDa. The expected band size for DNA Ligase IV/LIG4 is at 104 kDa.

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Figure 2. IF analysis of DNA Ligase IV/LIG4 using anti-DNA Ligase IV/LIG4 antibody (A01685-3) and anti-Tubulin Alpha antibody (M03989-3).
DNA Ligase IV/LIG4 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-DNA Ligase IV/LIG4 Antibody (A01685-3) and mouse anti-Tubulin Alpha antibody (M03989-3) 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 U937 cells using anti-DNA Ligase IV/LIG4 antibody (A01685-3).
Overlay histogram showing U937 cells stained with A01685-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNA Ligase IV/LIG4 Antibody (A01685-3, 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 LIG4 (Source: Uniprot.org, NCBI)

Alternative Names

DNA joinase; DNA ligase 4; DNA ligase IV; DNA repair enzyme; EC 6.5.1.1; ligase IV, DNA, ATP-dependent; Polydeoxyribonucleotide synthase [ATP] 4; polynucleotide ligase; sealase LIG4 LIG4S DNA ligase 4 DNA ligase 4|DNA joinase|DNA ligase IV|DNA repair enzyme|ligase IV, DNA, ATP-dependent|polydeoxyribonucleotide synthase [ATP] 4|polynucleotide ligase|sealase

For more info on LIG4, check out the LIG4 Infographic

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In this infographic, you will see the following information for LIG4: 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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