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Product Info Summary
| SKU: | A00544 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-SKP2 Antibody Picoband®
SKU/Catalog Number
A00544
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-SKP2 Antibody Picoband® catalog # A00544. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. 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-SKP2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00544)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human SKP2, which shares 88.9% and 94.4% amino acid (aa) sequence identity with mouse and rat SKP2, respectively.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00544 is reactive to SKP2 in Human, Mouse, Rat
Observed Molecular Weight
46, 48 kDa
Calculated molecular weight
47.8 kDa
Background of SKP2
The F box protein Skp2 (S-phase kinase-associated protein 2) is oncogenic, and its frequent amplification and overexpression correlate with the grade of malignancy of certain tumors. Skp2 controls p300-p53 signaling pathways in cancer cells, making it a potential molecular target for cancer therapy. This gene positively regulates the G (1)-S transition by controlling the stability of several G (1) regulators, such as the cell cycle inhibitor p27. This study provides evidence of a role for an F-box protein in oncogenesis and establishes SKP2 as a protooncogene causally involved in the pathogenesis of lymphomas.
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
A00544 is guaranteed for IF, 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, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Positive Control
WB: hhuman 293T whole cell, human Jurkat whole cell, human HepG2 whole cell, human MCF-7 whole cell, rat testis tissue, rat C6 whole cell, mouse testis tissue, mouse NIH/3T3 whole cell
IHC: human cervical cancer tissue
ICC/IF: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of SKP2 using anti-SKP2 antibody (A00544).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 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 Jurkat whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse testis tissue lysates,
Lane 8: mouse NIH/3T3 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-SKP2 antigen affinity purified polyclonal antibody (A00544) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for SKP2 at approximately 46, 48 kDa. The expected band size for SKP2 is at 46, 48 kDa.
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IHC analysis of SKP2 using anti-SKP2 antibody (A00544).
SKP2 was detected in a paraffin-embedded section of human cervical cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SKP2 Antibody (A00544) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of SKP2 using anti-SKP2 antibody (A00544) and anti-Tubulin Alpha antibody (M03989-3).
SKP2 was detected in immunocytochemical section of SiHa 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-SKP2 Antibody (A00544) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) 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.
Specific Publications For Anti-SKP2 Antibody Picoband® (A00544)
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