Rabbit Anti-SKP2 antibody | Sunlong Medical

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p45skp2; skp2p45; p45; skp2 p45;S-phase kinase-associated protein 2; SKP2; CDK2/Cyclin A associated protein p45; Cyclin A/CDK2 associated protein p45; F box protein Skp2; F box/LRR repeat protein 1; FBL 1; FBL1; FBXL 1; FBXL1; FLB 1; FLB1; MGC1366; SKP2_H

Cat:

SL1096R

Species Reactivity：

Human,Mouse,Rat,(predicted: Cow,Rabbit,)

Immunogen：

KLH conjugated synthetic peptide derived from human Skp2:351-424/424

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500ICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: 293T(Human) Cell Lysate at 40 ugPrimary: Anti-SKP2 (SL1096R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 48 kDObserved band size: 48 kDParaformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (SKP2) Polyclonal Antibody, Unconjugated (SL1096R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (SKP2) Polyclonal Antibody, Unconjugated (SL1096R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: human thyroid gland carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Skp2/skp2 p45 Polyclonal Antibody, Unconjugated(SL1096R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingHela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (SKP2) polyclonal Antibody, Unconjugated (SL1096R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Blank control: Hela(blue). Primary Antibody:Rabbit Anti- phospho-SKP2 antibody(SL1096R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions ); Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.ProtocolThe cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody (SL1096R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of SL1096R at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

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Unit:

Price: $220.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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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 ORC1L, 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. [SUBUNIT] Part of the SCF(SKP2) complex consisting of CUL1, RBX1, SKP1 and SKP2. Interacts directly with CUL1 and SKP1. Interacts with CKS1. Interacts with the cyclin A-CDK2 complex. Interacts with ORC1L, phosphorylated CDT1, phosphorylated RBL2, ELF4, phosphorylated RAG2, FOXO1A, UBP43, MYC, TOB1, TAL1 and MLL. [SIMILARITY] Contains 1 F-box domain. [SIMILARITY] Contains 8 LRR (leucine-rich) repeats.

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, FOXO1, UBP43, and probably MYC, TOB1 and TAL1. Degradation of TAL1 also requires STUB1. Recognizes CDKN1A in association with CCNE1 or CCNE2 and CDK2. Promotes ubiquitination and destruction of CDH1 in a CK1-Dependent Manner, thereby regulating cell migration.

Subunit:
Part of a SCF(SKP2) complex consisting of CUL1, RBX1, SKP1 and SKP2. Component of a SCF(SKP2)-like complex containing CUL1, SKP1, TRIM21 and SKP2. Interacts directly with CUL1 and SKP1. Interacts with CKS1. Interacts with the cyclin-A-CDK2 complex. Interacts with ORC1, phosphorylated CDT1, phosphorylated RBL2, ELF4, phosphorylated RAG2, FOXO1, UBP43, MYC, TOB1, TAL1 and MLL. Interacts with TRIM21.

Subcellular Location:
Cytoplasm. Nucleus.

Post-translational modifications:
Ubiquitinated by the APC/C complex, leading to its degradation by the proteasome. Deubiquitinated by USP13.
Acetylation at Lys-68 and Lys-71 increases stability through impairment of APC/SLCmediated proteolysis and promotes cytoplasmic retention. Deacetylated by SIRT3.

Similarity:
Contains 1 F-box domain.
Contains 9 LRR (leucine-rich) repeats.

SWISS:
Q13309

Gene ID:
6502

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

Unigene: 154278 Rat

Skp2异常表达可加速细胞周期转化，该蛋白与肿瘤分化程度有关，Skp2参与细胞转化和肿瘤的形成，目前主要用于消化系统肿瘤方面的研究。 Picture

Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (SKP2) Polyclonal Antibody, Unconjugated (SL1096R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (SKP2) Polyclonal Antibody, Unconjugated (SL1096R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: human thyroid gland carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Skp2/skp2 p45 Polyclonal Antibody, Unconjugated(SL1096R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (SKP2) polyclonal Antibody, Unconjugated (SL1096R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control: Hela(blue).
Primary Antibody:Rabbit Anti- phospho-SKP2 antibody(SL1096R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody (SL1096R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of SL1096R at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

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