Recombinant Anti-SKP2 antibody [EPR3305(2)] (ab183039)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3305(2)] to SKP2
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SKP2 antibody [EPR3305(2)]
See all SKP2 primary antibodies -
Description
Rabbit monoclonal [EPR3305(2)] to SKP2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: C6, NIH/3T3, F9, A549, MDA-MB-231, Jurkat, MCF7, HepG2, and NCCIT cell lysates. ICC: 293T cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3305(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab183039 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 |
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Flow Cyt (Intra) |
1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
1/100.
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/200. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
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Notes |
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Flow Cyt (Intra)
1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100. |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/200. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa). |
Target
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Function
Substrate 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. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Contains 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
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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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All lanes : Anti-SKP2 antibody [EPR3305(2)] (ab183039) at 1/200 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SKP2 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 48 kDaLanes 1 - 4: Merged signal (red and green). Green - ab183039 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab183039 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in SKP2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SKP2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab183039 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/200 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescenct analysis of 4% paraformaldehyde fixed 293T cells labeling SKP2 with ab183039 at 1/100 followed by Goat anti rabbit IgG(Alexa Fluor® 488) at 1/200 (green). Cells were counter stained with Dapi (blue).
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Intracellular flow cytometric analysis of 2% paraformaldehyde fixedHeLa cells labeling SKP2 with ab183039 at 1/70 followed byGoat anti rabbit IgG (FITC) at 1/150.Rabbit monoclonal IgG was used asIsotype control.
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All lanes : Anti-SKP2 antibody [EPR3305(2)] (ab183039) at 1/200 dilution
Lane 1 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates with 5% NFDM/TBST
Lane 5 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 6 : A549 (Human lung carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 7 : F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 8 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates with 5% NFDM/TBST
Lane 9 : C6 (Rat glial tumor glial cell) whole cell lysate with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Exposure time: 180 secondsWe recommend to increase the amount of samples or decrease antibody dilution to get clear bands.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
immunohistochemical analysis of paraffin embedded Human kidney clear cell carcinoma tissue labeling SKP2 with ab183039 at 1/50 followed by secondary staining with Ready to use HRP Polymer for Rabbit IgG and counterstained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (7)
ab183039 has been referenced in 7 publications.
- Han K & Zhou Q Skp2 Deteriorates the Uterine Receptivity by Interacting with HOXA10 and Promoting its Degradation. Reprod Sci 28:1069-1078 (2021). PubMed: 33104986
- Lv S et al. Downregulation of decidual SKP2 is associated with human recurrent miscarriage. Reprod Biol Endocrinol 19:88 (2021). PubMed: 34116705
- Wu D et al. A novel mitochondrial amidoxime reducing component 2 is a favorable indicator of cancer and suppresses the progression of hepatocellular carcinoma by regulating the expression of p27. Oncogene 39:6099-6112 (2020). PubMed: 32811980
- Yu C et al. Down-regulation of MTHFD2 inhibits NSCLC progression by suppressing cycle-related genes. J Cell Mol Med 24:1568-1577 (2020). PubMed: 31778025
- Liu D et al. 1,25-(OH)2D3/Vitamin D receptor alleviates systemic lupus erythematosus by downregulating Skp2 and upregulating p27. Cell Commun Signal 17:163 (2019). PubMed: 31823770
- Song J et al. Rottlerin promotes autophagy and apoptosis in gastric cancer cell lines. Mol Med Rep 18:2905-2913 (2018). PubMed: 30015872
- Dai Y et al. Activation of anaphase-promoting complex by p53 induces a state of dormancy in cancer cells against chemotherapeutic stress. Oncotarget 7:25478-92 (2016). WB . PubMed: 27009858