Anti-XRCC1 Antibody Picoband™ (monoclonal, 10E10)

Product Name

Anti-XRCC1 Antibody Picoband™ (monoclonal, 10E10)

View all XRCC1 Antibodies

SKU/Catalog Number

M00571-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-XRCC1 Antibody Picoband™ (monoclonal, 10E10) catalog # M00571-2. Tested in 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-XRCC1 Antibody Picoband™ (monoclonal, 10E10) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00571-2)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

10E10

Isotype

IgG2b

Immunogen

E. coli-derived human XRCC1 recombinant protein (Position: E538-A633).

*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

M00571-2 is reactive to XRCC1 in Human

Applications

M00571-2 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

95 kDa

Calculated molecular weight

69.477kDa

Background of XRCC1

XRCC1(X-RAY REPAIR, COMPLEMENTING DEFECTIVE, IN CHINESE HAMSTER, 1) is a DNA repair protein which complexes with DNA ligase III. The protein encoded by this gene is involved in the efficient repair of DNA single-strand breaks formed by exposure to ionizing radiation and alkylating agents. The XRCC1 gene is mapped to 19q13.31. The XRCC1 interacts with DNA ligase III, polymerase beta and poly (ADP-ribose) polymerase to participate in the base excision repair pathway. It may play a role in DNA processing during meiogenesis and recombination in germ cells. A rare microsatellite polymorphism in this gene is associated with cancer in patients of varying radiosensitivity. XRCC1 is phosphorylated in vivo and in vitro by CK2, and CK2 phosphorylation of XRCC1 on ser518, thr519, and thr523 largely determines aprataxin binding to XRCC1 through its FHA domain.

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

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of XRCC1 using anti-XRCC1 antibody (M00571-2).
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 Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Jurkat whole cell lysates,
Lane 5: human HEL whole cell lysates,
Lane 6: human A431 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 mouse anti-XRCC1 antigen affinity purified monoclonal antibody (Catalog # M00571-2) 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-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 XRCC1 at approximately 95 kDa. The expected band size for XRCC1 is at 69 kDa.

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Figure 2. IF analysis of XRCC1 using anti-XRCC1 antibody (M00571-2).
XRCC1 was detected in an immunocytochemical section of A431 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-XRCC1 Antibody (M00571-2) 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 Hela cells using anti-XRCC1 antibody (M00571-2).
Overlay histogram showing Hela cells stained with M00571-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-XRCC1 Antibody (M00571-2, 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 XRCC1 (Source: Uniprot.org, NCBI)

Alternative Names

DNA repair protein XRCC1; RCC; X-ray repair complementing defective repair in Chinese hamster cells 1; X-ray repair cross-complementing protein 1; X-ray-repair, complementing defective, repair in Chinese hamster XRCC1 RCC, SCAR26 X-ray repair cross complementing 1 DNA repair protein XRCC1|X-ray repair complementing defective repair in Chinese hamster cells 1|X-ray repair cross-complementing protein 1

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