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Product Info Summary
| SKU: | M00627 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC, ICC, WB |
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Product info
Product Name
Anti-APE1 APEX1 Antibody Picoband® (monoclonal, 5C11)
SKU/Catalog Number
M00627
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-APE1 APEX1 Antibody Picoband® (monoclonal, 5C11) catalog # M00627. Tested in Flow Cytometry, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-APE1 APEX1 Antibody Picoband® (monoclonal, 5C11) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00627)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
5C11
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human APE1 recombinant protein (Position: P2-L318). Human APE1 shares 94% and 93% amino acid (aa) sequence identity with mouse and rat APE1, respectively.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00627 is reactive to APEX1 in Human
Observed Molecular Weight
39 kDa
Calculated molecular weight
35.6 kDa
Background of APEX1
APEX1, also called apurinic endonuclease (APE), is a DNA repair enzyme having apurinic/apyrimidinic (AP) endonuclease, 3-prime, 5-prime-exonuclease, DNA 3-prime repair diesterase, and DNA 3-prime-phosphatase activities. The human APEX1 gene consists of 5 exons spanning 2.64 kb and exists as a single copy in the haploid genome. Using in situ hybridization, the APEX1 gene is mapped to 14q11.2-q12. The predicted APEX1 protein, which contained probable nuclear transport signals, was identified as a member of a family of DNA repair enzymes found in lower organisms. The abundance of the large form of APEX1 was increased in leiomyoma extracts relative to myometrial tissue extracts, and the large form was dominant in cell lines derived from leiomyosarcomas. The exonuclease activity of nuclear APEX1 can remove the anti-HIV nucleoside analogs AZT and D4T from the 3-prime terminus of a nick more efficiently than can cytosolic exonucleases.
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.
Application & Images
Applications
M00627 is guaranteed for Flow Cytometry, IHC, ICC, WB Boster Guarantee
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, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunohistochemistry (Frozen Section), 0.5-1μg/ml
Immunocytochemistry, 0.5-1μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human Hela whole cell, human MCF-7 whole cell, human COLO-320 whole cell, human HepG2 whole cell, Rabbit IgG, Marker 1113, human A549 whole cell, human PANC-1 whole cell, human 22RV1 whole cell, human MDA-MB-453 whole cell
IHC: human intestinal cancer tissue
FCM: SiHa cells
Validation Images & Assay Conditions
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Western blot analysis of APE1 using anti-APE1 antibody (M00627).
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 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human COLO-320 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: Rabbit IgG,
Lane 6: Marker 1113,
Lane 7: human A549 whole cell lysates ,
Lane 8: human PANC-1 whole cell lysates,
Lane 9: human 22RV1 whole cell lysates,
Lane 10: human MDA-MB-453 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-APE1 antigen affinity purified monoclonal antibody (Catalog # M00627) 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.
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IHC analysis of APE1 using anti-APE1 antibody (M00627).
APE1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-APE1 Antibody (M00627) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen. "
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Flow Cytometry analysis of SiHa cells using anti-APE1 antibody (M00627).
Overlay histogram showing SiHa cells stained with M00627 (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 mouse anti-APE1 Antibody (M00627, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-APE1 APEX1 Antibody Picoband® (monoclonal, 5C11) (M00627)
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