Anti-SKP2 antibody (ab68455)
- Datasheet
- References (6)
- Protocols
Overview
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Product nameAnti-SKP2 antibody
See all SKP2 primary antibodies -
DescriptionRabbit polyclonal to SKP2
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Host speciesRabbit
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Tested applicationsSuitable for: IP, WB, IHC-P, ICCmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide corresponding to Human SKP2 aa 2-20 (N terminal). Different from the related rat and mouse sequences by three amino acids.
Sequence:HRKHLQEIPDLSSNVATSF
Database link: Q13309
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferPreservatives: 0.025% Thimerosal (merthiolate), 0.025% Sodium azide
Constituents: 2.5% BSA, 0.45% Sodium chloride, 0.1% Dibasic monohydrogen sodium phosphate -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab68455 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP | Use a concentration of 5 µg/ml. | |
| WB | Use a concentration of 1 µg/ml. Predicted molecular weight: 48 kDa. | |
| IHC-P | Use a concentration of 1 - 2 µg/ml. | |
| ICC | Use a concentration of 1 - 2 µg/ml. Acetone fixed cells. Antigen retrieval by Pepsin and Trypsin is required. |
Target
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FunctionSubstrate recognition component of a SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins involved in cell cycle progression, signal transduction and transcription. Specifically recognizes phosphorylated CDKN1B/p27kip and is involved in regulation of G1/S transition. Degradation of CDKN1B/p27kip also requires CKS1. Recognizes target proteins ORC1, CDT1, RBL2, MLL, CDK9, RAG2, FOXO1A, UBP43, and probably MYC, TOB1 and TAL1. Degradation of TAL1 also requires STUB1. Recognizes CDKN1A in association with CCNE1 or CCNE2 and CDK2.
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PathwayProtein modification; protein ubiquitination.
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Sequence similaritiesContains 1 F-box domain.
Contains 9 LRR (leucine-rich) repeats. - Information by UniProt
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Database links
- Entrez Gene: 6502 Human
- Entrez Gene: 27401 Mouse
- Entrez Gene: 294790 Rat
- Omim: 601436 Human
- SwissProt: Q13309 Human
- SwissProt: Q9Z0Z3 Mouse
- Unigene: 23348 Human
- Unigene: 35584 Mouse
see all -
Alternative names
- CDK2/Cyclin A associated protein p45 antibody
- Cyclin A/CDK2 associated protein p45 antibody
- Cyclin-A/CDK2-associated protein p45 antibody
see all
Images
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Western blot - Anti-SKP2 antibody (ab68455)All lanes : Anti-SKP2 antibody (ab68455) at 1 µg/ml
Lane 1 : Whole cell lysates prepared from Hela cells
Lane 2 : Whole cell lysates prepared from 6T-CEM cells
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP conjugated Goat anti-rabbit IgG at 1/3000 dilution
Predicted band size: 48 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody (ab68455)ab68455 at 1µg/ml staining SKP2 in Human mammary cancer tissue sections by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). The tissue underwent enzymatic antigen retrieval in pepsin and trypsin. A Biotin-conjugated Goat anti-rabbit IgG was used as secondary at 1/200 dilution.
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Western blot - Anti-SKP2 antibody (ab68455)All lanes : Anti-SKP2 antibody (ab68455) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : Human placenta tissue lysate - total protein (ab29745)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?
Additional bands at: 29 kDa, 38 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
The predicted molecular weight of SKP2 is 48 kDa (SwissProt), however we expect to observe a banding pattern at 53 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. -
Immunoprecipitation - Anti-SKP2 antibody (ab68455)SKP2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to SKP2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab68455.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 53kDa; SKP2
Protocols
References
This product has been referenced in:
- Jang SM et al. The replication initiation determinant protein (RepID) modulates replication by recruiting CUL4 to chromatin. Nat Commun 9:2782 (2018). Read more (PubMed: 30018425) »
- Ma X et al. Ubiquitin conjugating enzyme E2 L3 promoted tumor growth of NSCLC through accelerating p27kip1 ubiquitination and degradation. Oncotarget 8:84193-84203 (2017). Read more (PubMed: 29137415) »
