Product: CDC25C Antibody
Catalog: AF6258
Description: Rabbit polyclonal antibody to CDC25C
Application: WB IHC IF/ICC
Reactivity: Human, Mouse
Prediction: Horse, Dog, Xenopus
Mol.Wt.: 60kDa; 53kD(Calculated).
Uniprot: P30307
RRID: AB_2835119

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, 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
Prediction:
Horse(90%), Dog(90%), Xenopus(90%)
Clonality:
Polyclonal
Specificity:
CDC25C Antibody detects endogenous levels of total CDC25C.
RRID:
AB_2835119
Cite Format: Affinity Biosciences Cat# AF6258, RRID:AB_2835119.
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

CDC 25; Cdc 25C; CDC25; CDC25C; Cell division cycle 25 homolog C; Cell division cycle 25C; Cell division cycle 25C protein; Dual specificity phosphatase Cdc25C; M phase inducer phosphatase 3; M-phase inducer phosphatase 3; Mitosis inducer CDC25; MPIP3; MPIP3_HUMAN; Phosphotyrosine phosphatase; PPP1R60; protein phosphatase 1, regulatory subunit 60;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Description:
a member of the MPI phosphatase family. Activates the cyclin dependent kinase CDC2 by removing two phosphate groups and it is required for entry into mitosis. Shuttles between the nucleus and the cytoplasm due to nuclear localization and nuclear export signals.
Sequence:
MSTELFSSTREEGSSGSGPSFRSNQRKMLNLLLERDTSFTVCPDVPRTPVGKFLGDSANLSILSGGTPKRCLDLSNLSSGEITATQLTTSADLDETGHLDSSGLQEVHLAGMNHDQHLMKCSPAQLLCSTPNGLDRGHRKRDAMCSSSANKENDNGNLVDSEMKYLGSPITTVPKLDKNPNLGEDQAEEISDELMEFSLKDQEAKVSRSGLYRSPSMPENLNRPRLKQVEKFKDNTIPDKVKKKYFSGQGKLRKGLCLKKTVSLCDITITQMLEEDSNQGHLIGDFSKVCALPTVSGKHQDLKYVNPETVAALLSGKFQGLIEKFYVIDCRYPYEYLGGHIQGALNLYSQEELFNFFLKKPIVPLDTQKRIIIVFHCEFSSERGPRMCRCLREEDRSLNQYPALYYPELYILKGGYRDFFPEYMELCEPQSYCPMHHQDHKTELLRCRSQSKVQEGERQLREQIALLVKDMSP

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
Horse
90
Dog
90
Xenopus
90
Pig
60
Bovine
60
Sheep
60
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P30307 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S14 Phosphorylation
S15 Phosphorylation
S17 Phosphorylation
S20 Phosphorylation
K27 Ubiquitination
T37 Phosphorylation
S38 Phosphorylation
T48 Phosphorylation P06493 (CDK1)
K52 Ubiquitination
S57 Phosphorylation
S61 Phosphorylation
S64 Phosphorylation
T67 Phosphorylation P06493 (CDK1)
K69 Ubiquitination
S75 Phosphorylation
S122 Phosphorylation P06493 (CDK1)
S129 Phosphorylation
T130 Phosphorylation P06493 (CDK1)
S146 Phosphorylation
S148 Phosphorylation
K151 Ubiquitination
Y165 Phosphorylation
S168 Phosphorylation P45984 (MAPK9) , P53779 (MAPK10) , P45983 (MAPK8) , P06493 (CDK1)
T171 Phosphorylation
T172 Phosphorylation
S191 Phosphorylation Q9H4B4 (PLK3)
S198 Phosphorylation Q9H4B4 (PLK3) , PR:P53350 (hPLK1) , P53350 (PLK1)
S207 Phosphorylation P53778 (MAPK12)
S209 Phosphorylation
Y212 Phosphorylation
S214 Phosphorylation P06493 (CDK1) , P24941 (CDK2)
S216 Phosphorylation P27361 (MAPK3) , Q13237 (PRKG2) , Q13535 (ATR) , Q683Z8 (CHK2) , P28482 (MAPK1) , Q16539 (MAPK14) , P49137 (MAPKAPK2) , Q16512 (PKN1) , O14757 (CHEK1) , Q9H4B4 (PLK3) , P27448 (MARK3) , P43405 (SYK) , Q8TDC3 (BRSK1) , O96017 (CHEK2)
K233 Ubiquitination
T236 Phosphorylation P68400 (CSNK2A1)
K240 Ubiquitination
K244 Ubiquitination
S247 Phosphorylation Q15418 (RPS6KA1)
K251 Ubiquitination
K260 Ubiquitination
S263 Phosphorylation
S287 Phosphorylation
K288 Ubiquitination
K298 Ubiquitination
K303 Ubiquitination
K317 Ubiquitination
K360 Ubiquitination
Y401 Phosphorylation
K413 Ubiquitination
Y416 Phosphorylation
S449 Phosphorylation
K452 Ubiquitination
K469 Ubiquitination
S472 Phosphorylation

Research Backgrounds

Function:

Functions as a dosage-dependent inducer in mitotic control. Tyrosine protein phosphatase required for progression of the cell cycle. When phosphorylated, highly effective in activating G2 cells into prophase. Directly dephosphorylates CDK1 and activates its kinase activity.

PTMs:

Phosphorylated by CHEK1 and MAPK14 at Ser-216. This phosphorylation creates a binding site for 14-3-3 protein and inhibits the phosphatase. Phosphorylated by PLK4. Phosphorylated by PLK1, leading to activate the phosphatase activity. Phosphorylation by PLK3 at Ser-191 promotes nuclear translocation. Ser-198 is a minor phosphorylation site. Was initially reported to be phosphorylated by PLK3 at Ser-216. However, such phosphorylation by PLK3 was not confirmed by other groups. Phosphorylation at Thr-48, Thr-67, Ser-122, Thr-130, Ser-168 and Ser-214 occurs at G2 and G2-M transition and is probably catalyzed by CDK1. Ser-168 phosphorylation levels are lower than those at the other 5 CDK1 sites. Phosphorylation by CDK1 leads to increased activity.

Subcellular Location:

Nucleus.

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

Interacts with MAPK14 and 14-3-3 proteins. When phosphorylated on Ser-129 and/or Thr-130, interacts with PLK1.

(Microbial infection) Interacts with HIV-1 Vpr; this interaction inactivates CDC25C phosphatase activity.

Family&Domains:

Belongs to the MPI phosphatase family.

Research Fields

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

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

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

· Organismal Systems > Endocrine system > Progesterone-mediated oocyte maturation.

References

1). Xue T et al. Cyclovirobuxine D inhibits growth and progression of non‑small cell lung cancer cells by suppressing the KIF11‑CDC25C‑CDK1‑CyclinB1 G2/M phase transition regulatory network and the NFκB/JNK signaling pathway. International Journal of Oncology 2023 May;62(5) (PubMed: 36929198) [IF=5.2]

Application: WB    Species: Human    Sample: NSCLC cells

Figure 6 KIF11, KIF11-CDK1-CDC25C-cyclinB1 oncogenic network and NF-kB/JNK signaling pathway are the potential CVB-D therapeutic targets in the NSCLC cells. (A) Venn diagram showed the overlap number of potential CVB-D target genes in NSCLC cells by intersecting four LUAD datasets. (B) Protein-protein interaction network analysis revealed interaction between the 10 overlapping DEGs. The disconnected nodes are hidden. (C) The interactive heatmap from GEPIA 2 database showed expression of the 6 DEGs in LUAD and normal lung tissues. KIF11 is the most differentially expressed gene between LUAD and normal lung tissues. (D) Venn diagram revealed 2 common genes (KIF11 and CDK1) based on intersection between the 10 overlapping DEGs and the 100 predicted CVB-D target genes extracted from the Swisstarget prediction database. (E) Western blot analysis identified the expression of KIF11, CDK1, CDC25C and cyclinB1 proteins in the control and CVB-D-treated NSCLC cells. (F and G) Immunohistochemical assay results showed the expression levels and localization of the KIF11 protein in the control and CVB-D-treated A549 xenograft tumor tissues. (H) Western blot assay results revealed the KIF11 protein levels in the si-NC and si-KIF11-transfected NSCLC cells. (I) Western blot analysis demonstrated the expression levels of the oncogenic signaling network proteins (KIF11, CDK1, CDC25C and CyclinB1) and the NF-κB/JNK signaling pathway proteins (p65, p-p65, JNK and p-JNK) in the si-NC- and si-KIF11-transfected NSCLC cells. **P

Application: IHC    Species: Mouse    Sample: A549 cells

Figure 7 CVB-D inhibits in vivo progression of A549 xenografts tumors. (A) Experimental design for tumor xenograft experiments of NSCLC in nude mice to evaluate the in vivo therapeutic effects of CVB-D. Specific time points for CVB-D treatment and experimental analysis are indicated. (B) Representative images of the NSCLC xenograft tumor models with or without CVB-D treatment. (C) Representative images of the NSCLC xenograft tumor tissues from control and CVB-D-treated mice. (D and E) NSCLC xenograft tumor weights and volumes in the control and CVB-D treatment groups of nude mice. (F) Representative H&E and TUNEL staining images of the A549 xenograft tumor sections in the control and CVB-D treatment groups of nude mice (magnification, ×200; scale bar, 50 µm). (G) The proportion of apoptotic cells in the A549 xenograft tumors derived from the control and CVB-D-treatment groups of nude mice based on TUNEL staining. (H) The body weights of the xenograft tumor model mice in the control and CVB-D treatment groups. (I and J) Immunohistochemical assay data revealed the expression levels and distribution of CDK1, CDC25C, cyclinB1, Ki67, Bcl-2 and N-cadherin proteins in the A549 xenograft tumors derived from the control and CVB-D treatment groups (magnification, ×200; scale bar, 50 µm). *P

2). Bai Y et al. BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo. MOLECULAR CARCINOGENESIS 2017 Mar;56(3):1137-1149 (PubMed: 27770580) [IF=4.6]

Application: WB    Species: human    Sample:

(D) Effects of BCL2L10 on protein expression of PCNA and CDC25C by Western blot.

3). Wang Y et al. Ochratoxin A induces cytotoxicity through ROS-mediated endoplasmic reticulum stress pathway in human gastric epithelium cells. TOXICOLOGY 2022 Sep 1;479:153309. (PubMed: 36058351) [IF=4.5]

4). Li AD et al. TGR5 promotes cholangiocarcinoma by interacting with mortalin. EXPERIMENTAL CELL RESEARCH 2020 Jan 21:111855 (PubMed: 31978385) [IF=3.7]

Application: WB    Species: Human    Sample: CC cell

Fig. 2. The elevated TGR5 expression promotes the proliferation of the CC cell line. (A) The Western blot analysis indicated that TGR5 expession was successfully regulated with the TGR5 recombinant plasmid and siRNA targeting TGR5 in RBE cells and QBC-939 cells. The upregulation of TGR5 induced elevated expression of proliferating cell nuclear antigen (PCNA), which is involved in DNA replication-linked processes, and is a known molecular marker of proliferation, whereas silencing of TGR5 lead to a reduced expression of PCNA. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as a normalizing control. (B) CC cells treated with TGR5 siRNA revealed a weaker cell viability while these treated with TGR5 recombinant plasmid exhibited an increased cell viability, as have shown by CCK-8 assay, and this was most pronouncing at 72 h point. (C) The BrdU incorporation test revealed that the DNA synthesis in CC cells was accelerated with the TGR5 recombinant plasmid and was repressed with siRNA targeting TGR5.(D) CDC25C and cyclin D1 play a crucial role in accelerating cell cycle procession. The exposure to the TGR5 plasmid led to the augmented expression of CDC25C in RBE and QBC-939 cells, and increase the expression of cyclin D1 in QBC-939 cells. *P < 0.05 and **P < 0.01, when compared with the control group. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as a normalizing control.

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