Product: p27 Kip1 Antibody
Catalog: AF6324
Description: Rabbit polyclonal antibody to p27 Kip1
Application: WB IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 27kDa; 22kD(Calculated).
Uniprot: P46527
RRID: AB_2835182

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(90%)
Clonality:
Polyclonal
Specificity:
p27 Kip1 Antibody detects endogenous levels of total p27 Kip1.
RRID:
AB_2835182
Cite Format: Affinity Biosciences Cat# AF6324, RRID:AB_2835182.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AA408329; AI843786; Cdki1b; CDKN 1B; CDKN 4; CDKN1B; CDKN4; CDN1B_HUMAN; Cyclin Dependent Kinase Inhibitor 1B; Cyclin dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor 1B (p27, Kip1); Cyclin-dependent kinase inhibitor 1B; Cyclin-dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor p27 Kip1; KIP 1; KIP1; MEN1B; MEN4; OTTHUMP00000195098; OTTHUMP00000195099; p27; p27 Kip1; P27-like cyclin-dependent kinase inhibitor; p27Kip1;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P46527 CDN1B_HUMAN:

Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.

Description:
This gene encodes a cyclin-dependent kinase inhibitor, which shares a limited similarity with CDK inhibitor CDKN1A/p21. The encoded protein binds to and prevents the activation of cyclin E-CDK2 or cyclin D-CDK4 complexes, and thus controls the cell cycle progression at G1.
Sequence:
MSNVRVSNGSPSLERMDARQAEHPKPSACRNLFGPVDHEELTRDLEKHCRDMEEASQRKWNFDFQNHKPLEGKYEWQEVEKGSLPEFYYRPPRPPKGACKVPAQESQDVSGSRPAAPLIGAPANSEDTHLVDPKTDPSDSQTGLAEQCAGIRKRPATDDSSTQNKRANRTEENVSDGSPNAGSVEQTPKKPGLRRRQT

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Chicken
90
Pig
0
Horse
0
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P46527 As Substrate

Site PTM Type Enzyme
S2 O-Glycosylation
S10 Phosphorylation Q9UQM7 (CAMK2A) , Q00534 (CDK6) , Q9H2X6 (HIPK2) , Q8TAS1 (UHMK1) , Q00536 (CDK16) , P31749 (AKT1) , P28482 (MAPK1) , Q9Y463 (DYRK1B)
S12 Phosphorylation
R15 Methylation
K25 Ubiquitination
K68 Ubiquitination
Y74 Phosphorylation P07947 (YES1) , P12931 (SRC)
K81 Acetylation
K81 Ubiquitination
S83 Phosphorylation P68400 (CSNK2A1)
Y88 Phosphorylation O60674 (JAK2) , P12931 (SRC) , P00519 (ABL1) , P07947 (YES1) , P07948 (LYN)
Y89 Phosphorylation P12931 (SRC) , P00519 (ABL1) , P07947 (YES1)
S106 O-Glycosylation
S110 O-Glycosylation
K134 Ubiquitination
S140 Phosphorylation Q13315 (ATM)
K153 Ubiquitination
R154 Methylation
T157 O-Glycosylation
T157 Phosphorylation Q14012 (CAMK1) , P31749 (AKT1) , P31751 (AKT2) , P11309-2 (PIM1) , O00141 (SGK1) , Q86V86 (PIM3) , Q9P1W9 (PIM2)
K165 Ubiquitination
S175 Phosphorylation
S178 Phosphorylation P28482 (MAPK1)
S183 Phosphorylation
T187 Phosphorylation Q00534 (CDK6) , P31749 (AKT1) , P28482 (MAPK1) , Q00535 (CDK5) , P27361 (MAPK3) , P24941 (CDK2)
T198 O-Glycosylation
T198 Phosphorylation P11309-2 (PIM1) , P31749 (AKT1) , Q14012 (CAMK1) , P51812 (RPS6KA3) , Q13131 (PRKAA1) , Q86V86 (PIM3) , Q15418 (RPS6KA1) , O00141 (SGK1) , Q9P1W9 (PIM2)

Research Backgrounds

Function:

Important regulator of cell cycle progression. Inhibits the kinase activity of CDK2 bound to cyclin A, but has little inhibitory activity on CDK2 bound to SPDYA. Involved in G1 arrest. Potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. Forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. Acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry.

PTMs:

Phosphorylated; phosphorylation occurs on serine, threonine and tyrosine residues. Phosphorylation on Ser-10 is the major site of phosphorylation in resting cells, takes place at the G(0)-G(1) phase and leads to protein stability. Phosphorylation on other sites is greatly enhanced by mitogens, growth factors, cMYC and in certain cancer cell lines. The phosphorylated form found in the cytoplasm is inactivate. Phosphorylation on Thr-198 is required for interaction with 14-3-3 proteins. Phosphorylation on Thr-187, by CDK1 and CDK2 leads to protein ubiquitination and proteasomal degradation. Tyrosine phosphorylation promotes this process. Phosphorylation by PKB/AKT1 can be suppressed by LY294002, an inhibitor of the catalytic subunit of PI3K. Phosphorylation on Tyr-88 and Tyr-89 has no effect on binding CDK2, but is required for binding CDK4. Dephosphorylated on tyrosine residues by G-CSF.

Ubiquitinated; in the cytoplasm by the KPC complex (composed of RNF123/KPC1 and UBAC1/KPC2) and, in the nucleus, by SCF(SKP2). The latter requires prior phosphorylation on Thr-187. Ubiquitinated; by a TRIM21-containing SCF(SKP2)-like complex; leads to its degradation.

Subject to degradation in the lysosome. Interaction with SNX6 promotes lysosomal degradation (By similarity).

Subcellular Location:

Nucleus. Cytoplasm. Endosome.
Note: Nuclear and cytoplasmic in quiescent cells. AKT- or RSK-mediated phosphorylation on Thr-198, binds 14-3-3, translocates to the cytoplasm and promotes cell cycle progression. Mitogen-activated UHMK1 phosphorylation on Ser-10 also results in translocation to the cytoplasm and cell cycle progression. Phosphorylation on Ser-10 facilitates nuclear export. Translocates to the nucleus on phosphorylation of Tyr-88 and Tyr-89. Colocalizes at the endosome with SNX6; this leads to lysosomal degradation (By similarity).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.

Subunit Structure:

Forms a ternary complex composed of CCNE1, CDK2 and CDKN1B. Interacts directly with CCNE1; the interaction is inhibited by CDK2-dependent phosphorylation on Thr-187. Interacts with COPS5, subunit of the COP9 signalosome complex; the interaction leads to CDKN1B degradation. Interacts with NUP50; the interaction leads to nuclear import and degradation of phosphorylated CDKN1B. Interacts with CCND1 and SNX6 (By similarity). Interacts (Thr-198-phosphorylated form) with 14-3-3 proteins, binds strongly YWHAQ, weakly YWHAE and YWHAH, but not YWHAB nor YWHAZ; the interaction with YWHAQ results in translocation to the cytoplasm. Interacts with AKT1 and LYN; the interactions lead to cytoplasmic mislocation, phosphorylation of CDKN1B and inhibition of cell cycle arrest. Forms a ternary complex with CCNA2 and CDK2; CDKN1B inhibits the kinase activity of CDK2 through conformational rearrangements. Interacts (unphosphorylated form) with CDK2. Forms a complex with CDK2 and SPDYA, but does not directly interact with SPDYA. Forms a ternary complex composed of cyclin D, CDK4 and CDKN1B. Interacts (phosphorylated on Tyr-88 and Tyr-89) with CDK4; the interaction is required for cyclin D and CDK4 complex assembly, induces nuclear translocation and activates the CDK4 kinase activity. Interacts with GRB2. Interacts with PIM1. Identified in a complex with SKP1, SKP2 and CKS1B. Interacts with UHMK1; the interaction leads to cytoplasmic mislocation, phosphorylation of CDKN1B and inhibition of cell cycle arrest. Interacts also with CDK1. Dephosphorylated on Thr-187 by PPM1H, leading to CDKN1B stability.

Family&Domains:

A peptide sequence containing only AA 28-79 retains substantial Kip1 cyclin A/CDK2 inhibitory activity.

Belongs to the CDI family.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

References

1). Wu J et al. Skp2 modulates proliferation, senescence and tumorigenesis of glioma. Cancer Cell International 2020 Mar 6;20:71. (PubMed: 32165861) [IF=5.8]

Application: WB    Species: mouse    Sample: U87, U138, and LNZ308 cells

Fig. 2| Downregulation of Skp2 attenuated in vitro and in vivo cell proliferation. a On Western blot analyses, the protein level of Skp2 in glioma cell lines (A172, U87, U118, U373, LNZ308, U138 and U343), as compared to normal astrocytes (AST), was enhanced. b Skp2 was successfully knocked-down by two shRNA fragments in U87, U138, and LNZ308 cells compared with the negative control shLuc. The protein levels of the downstream molecules ­p21Cip1/Waf1 and ­p27Kip1 were increased upon the knockdown of Skp2 in all three cell lines.

2). Huang H et al. Ligustrazine Suppresses Platelet-Derived Growth Factor-BB-Induced Pulmonary Artery Smooth Muscle Cell Proliferation and Inflammation by Regulating the PI3K/AKT Signaling Pathway. The American journal of Chinese medicine 2021 Feb 20;1-23. (PubMed: 33622214) [IF=5.7]

3). Geng QS et al. Over-Expression and Prognostic Significance of FATP5, as a New Biomarker, in Colorectal Carcinoma. Frontiers in Molecular Biosciences 2022 Jan 27;8:770624. (PubMed: 35155561) [IF=5.0]

4). Lin Y et al. LncRNA GACAT3 acts as a competing endogenous RNA of HMGA1 and alleviates cucurbitacin B-induced apoptosis of gastric cancer cells. GENE 2018 Aug 8 (PubMed: 30098426) [IF=3.5]

Application: WB    Species: human    Sample: SGC-7901 cells

Fig. 3. |GACAT3 and HMGA1 promote cell growth by inhibiting p21 and p27.qRT-PCR and western blot analyses of p27 and p21 levels in gastric cancer cells after GACAT3 or HMGA1 siRNA-mediated knockdown. The mRNA expression levels of p27 and p21 in SGC-7901 (A) and BGC-823 (B) cells. Western blotting results and relative protein levels of p27, p21, and HMGA1 in SGC-7901 (C) and BGC-823 (D)cells. Data represent the mean ± S.D. of three biological replicates. *p < 0.05; **p < 0.01.

5). Wang X et al. Regulation of the PTEN/PI3K/AKT pathway in RCC using the active compounds of natural products in vitro. Molecular Medicine Reports 2021 Nov;24(5):766. (PubMed: 34490473) [IF=3.4]

Application: WB    Species: Human    Sample: RCC cells

Figure 3. Naringenin arrests cell cycle progression of renal cell carcinoma cells in the G2 phase. 786-O and OS-RC-2 cells were treated with naringenin (0, 4 or 8 µM) for 48 h. (A) Cell cycle distribution was determined using flow cytometry. (B) Western blot analysis of cyclin E1, cyclin A2, cyclin B1, cyclin D1, P27 and P21 expression levels. Data are presented as the mean ± SD. *P<0.05, **P<0.01 vs. Abs. Abs, absolute ethanol.

Application: WB    Species: human    Sample: 786‑O and OS‑RC‑2 cells

Figure 3.| Naringenin arrests cell cycle progression of renal cell carcinoma cells in the G2 phase. 786‑O and OS‑RC‑2 cells were treated with naringenin (0, 4 or 8 µM) for 48 h. (A) Cell cycle distribution was determined using flow cytometry. (B) Western blot analysis of cyclin E1, cyclin A2, cyclin B1, cyclin D1, P27 and P21 expression levels. Data are presented as the mean ± SD. * P<0.05, **P<0.01 vs. Abs. Abs, absolute ethanol.

6). Geng F et al. Fusobacterium nucleatum Caused DNA Damage and Promoted Cell Proliferation by the Ku70/p53 Pathway in Oral Cancer Cells. DNA AND CELL BIOLOGY 2019 Nov 25 (PubMed: 31765243) [IF=3.1]

Application: WB    Species: human    Sample: Tca8113 tongue squamous cell

FIG. 3.| The protein expression of p27, wild p53, and Ku70 in response to F. nucleatum infection (A).

7). Liang R et al. Dihydroartemisinin inhibits the tumorigenesis and invasion of gastric cancer by regulating STAT1/KDR/MMP9 and P53/BCL2L1/CASP3/7 pathways. PATHOLOGY RESEARCH AND PRACTICE 2020 Dec 13;218:153318. (PubMed: 33370709) [IF=2.8]

Application: WB    Species: mice    Sample: GC cells

Fig. 6. DHA regulated p53/BCL2L1/Caspase-3/-7 and STAT1/KDR/MMP9 signaling pathways. (A) qRT-PCR analysis of the effects of DHA on the expression levels of BCL2, BAX, CKD4, p53/BCL2L1/Caspase-3/-7 and STAT1/KDR/MMP9 in GC cells. (B) WB analysis of the effects of DHA on the expression of apoptosis and cycle-related proteins including cleaved-caspase 3, caspase 3, caspase 7, BCL2L1, p27 and p53. (C) WB analysis of the effects of DHA on the expression of p-STAT1, STAT1, p-KDR, KDR and MMP9.

8). Chen J et al. LINC00152 acts as a competing endogenous RNA of HMGA1 to promote the growth of gastric cancer cells. JOURNAL OF CLINICAL LABORATORY ANALYSIS 2022 Feb;36(2):e24192. (PubMed: 35014092) [IF=2.7]

Application: WB    Species: Human    Sample: GC cells

FIGURE 4 HMGA1 and LINC00152 regulate cell cycle progression. (A) The relative expression of P27 was measured after siRNA transfection. (B) Western blot analysis of HMGA1, P27, and β‐actin 48 h after siRNA transfection. (C) The relative HMGA1 and P27 protein levels were quantified from B. Data represent the mean ± S.D. (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001

9). Zhang Q et al. S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors. American Journal of Translational Research 2018 Mar 15;10(3):731-743 (PubMed: 29636863) [IF=2.2]

Application: WB    Species: human    Sample: 5637 and T24 cells

Figure 4.| miR-1236 induced Skp2 expression independent of p21 activation and influenced downstream Skp2 gene expression. F. P27 and CDK1 expression levels were detected by Western blot in 5637 and T24 cells. miR-1236 decreased p27 expression and up-regulated CDK1 expression. dsRNA-245 had no significant effect on p27 and CDK1.

10). Yang Q et al. Methyl-β-cyclodextrin potentiates the BITC-induced anti-cancer effect through modulation of the Akt phosphorylation in human colorectal cancer cells. BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 2018 Dec;82(12):2158-2167 (PubMed: 30200817) [IF=1.6]

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