CDC25A Antibody - #AF6252

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Product: CDC25A Antibody
Catalog: AF6252
Description: Rabbit polyclonal antibody to CDC25A
Application: WB IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 60kDa; 59kD(Calculated).
Uniprot: P30304
RRID: AB_2835116

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 100ul $280 In stock
 200ul $350 In stock

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

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:
Pig(100%), Bovine(100%), Horse(100%), Sheep(91%), Rabbit(100%), Dog(91%), Chicken(80%)
Clonality:
Polyclonal
Specificity:
CDC25A Antibody detects endogenous levels of total CDC25A.
RRID:
AB_2835116
Cite Format: Affinity Biosciences Cat# AF6252, RRID:AB_2835116.
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 25a; CDC25A; CDC25A2; CDC25A2 CAG isoform; Cell division cycle 25 homolog A (S. pombe); Cell division cycle 25A; Cell division cycle 25A isoform a; Cell division cycle 25A isoform b; D9Ertd393e; Dual specificity phosphatase Cdc25A; M phase inducer phosphatase 1; M-phase inducer phosphatase 1; MGC115549; MPIP1_HUMAN;

Immunogens

Immunogen:
Uniprot:

P30304(MPIP1_HUMAN) >>Visit HPA database.

Gene(ID):
CDC25A(993)
Description:
CDC25A is a member of the CDC25 family of phosphatases. CDC25A is required for progression from G1 to the S phase of the cell cycle. It activates the cyclin-dependent kinase CDC2 by removing two phosphate groups.
Sequence:
MELGPEPPHRRRLLFACSPPPASQPVVKALFGASAAGGLSPVTNLTVTMDQLQGLGSDYEQPLEVKNNSNLQRMGSSESTDSGFCLDSPGPLDSKENLENPMRRIHSLPQKLLGCSPALKRSHSDSLDHDIFQLIDPDENKENEAFEFKKPVRPVSRGCLHSHGLQEGKDLFTQRQNSAPARMLSSNERDSSEPGNFIPLFTPQSPVTATLSDEDDGFVDLLDGENLKNEEETPSCMASLWTAPLVMRTTNLDNRCKLFDSPSLCSSSTRSVLKRPERSQEESPPGSTKRRKSMSGASPKESTNPEKAHETLHQSLSLASSPKGTIENILDNDPRDLIGDFSKGYLFHTVAGKHQDLKYISPEIMASVLNGKFANLIKEFVIIDCRYPYEYEGGHIKGAVNLHMEEEVEDFLLKKPIVPTDGKRVIVVFHCEFSSERGPRMCRYVRERDRLGNEYPKLHYPELYVLKGGYKEFFMKCQSYCEPPSYRPMHHEDFKEDLKKFRTKSRTWAGEKSKREMYSRLKKL

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
Pig
100
Horse
100
Bovine
100
Rabbit
100
Sheep
91
Dog
91
Chicken
80
Xenopus
60
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P30304 As Substrate

Site PTM Type Enzyme
Ubiquitination
S18 Phosphorylation P06493 (CDK1)
S40 Phosphorylation
Y59 Phosphorylation
S76 Phosphorylation O14757 (CHEK1) , P49841 (GSK3B)
S77 Phosphorylation
S79 Phosphorylation Q8NG66 (NEK11) , P48729 (CSNK1A1)
T80 Phosphorylation Q9H4B4 (PLK3)
S82 Phosphorylation P48729 (CSNK1A1) , Q8NG66 (NEK11)
S88 Phosphorylation Q8NG66 (NEK11)
S107 Phosphorylation
K111 Ubiquitination
S116 Phosphorylation P06493 (CDK1)
K120 Ubiquitination
S124 Phosphorylation Q683Z8 (CHK2) , O96017 (CHEK2) , O14757 (CHEK1)
S126 Phosphorylation
K150 Acetylation
K150 Ubiquitination
K169 Ubiquitination
S178 Phosphorylation Q683Z8 (CHK2) , O14757 (CHEK1) , O96017 (CHEK2)
K257 Ubiquitination
S261 Phosphorylation
S263 Phosphorylation P24941 (CDK2)
R278 Methylation
S279 Phosphorylation Q683Z8 (CHK2) , O14757 (CHEK1) , O96017 (CHEK2)
S283 Phosphorylation
R290 Methylation
S293 Phosphorylation O14757 (CHEK1) , O96017 (CHEK2) , Q15418 (RPS6KA1) , Q683Z8 (CHK2)
S295 Phosphorylation Q15418 (RPS6KA1)
K300 Ubiquitination
S321 Phosphorylation
K343 Ubiquitination
K353 Ubiquitination
K358 Ubiquitination
Y359 Phosphorylation
S361 Phosphorylation
S367 Phosphorylation
K372 Ubiquitination
K378 Ubiquitination
K415 Ubiquitination
K471 Ubiquitination
Y486 Phosphorylation
T507 Phosphorylation O14757 (CHEK1)
S513 Phosphorylation Q9H4B4 (PLK3)
Y518 Phosphorylation
S519 Phosphorylation Q9H4B4 (PLK3)

Research Backgrounds

Function:

Tyrosine protein phosphatase which functions as a dosage-dependent inducer of mitotic progression. Directly dephosphorylates CDK1 and stimulates its kinase activity. Also dephosphorylates CDK2 in complex with cyclin E, in vitro.

PTMs:

Phosphorylated by CHEK1 on Ser-76, Ser-124, Ser-178, Ser-279, Ser-293 and Thr-507 during checkpoint mediated cell cycle arrest. Also phosphorylated by CHEK2 on Ser-124, Ser-279, and Ser-293 during checkpoint mediated cell cycle arrest. Phosphorylation on Ser-178 and Thr-507 creates binding sites for YWHAE/14-3-3 epsilon which inhibits CDC25A. Phosphorylation on Ser-76, Ser-124, Ser-178, Ser-279 and Ser-293 may also promote ubiquitin-dependent proteolysis of CDC25A by the SCF complex. Phosphorylation of CDC25A at Ser-76 by CHEK1 primes it for subsequent phosphorylation at Ser-79, Ser-82 and Ser-88 by NEK11. Phosphorylation by NEK11 is required for BTRC-mediated polyubiquitination and degradation. Phosphorylation by PIM1 leads to an increase in phosphatase activity. Phosphorylated by PLK3 following DNA damage, leading to promote its ubiquitination and degradation.

Ubiquitinated by the anaphase promoting complex/cyclosome (APC/C) ubiquitin ligase complex that contains FZR1/CDH1 during G1 phase leading to its degradation by the proteasome. Ubiquitinated by a SCF complex containing BTRC and FBXW11 during S phase leading to its degradation by the proteasome. Deubiquitination by USP17L2/DUB3 leads to its stabilization.

Subunit Structure:

Interacts with CCNB1/cyclin B1. Interacts with YWHAE/14-3-3 epsilon when phosphorylated. Interacts with CUL1 specifically when CUL1 is neddylated and active. Interacts with BTRC/BTRCP1 and FBXW11/BTRCP2. Interactions with CUL1, BTRC and FBXW11 are enhanced upon DNA damage. Interacts with PIM1. Interacts with CHEK2; mediates CDC25A phosphorylation and degradation in response to infrared-induced DNA damages. Interacts with HSP90AB1; prevents heat shock-mediated CDC25A degradation and contributes to cell cycle progression.

Family&Domains:

The phosphodegron motif mediates interaction with specific F-box proteins when phosphorylated. Putative phosphorylation sites at Ser-79 and Ser-82 appear to be essential for this interaction.

Belongs to the MPI phosphatase family.

Research Fields

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

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

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

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

References

1). Novel LncRNA OXCT1-AS1 indicates poor prognosis and contributes to tumorigenesis by regulating miR-195/CDC25A axis in glioblastoma. Journal of Experimental & Clinical Cancer Research (PubMed: 33832517) [IF=11.3]
2). Inhibition of SGLT1 protects against glycemic variability-induced cardiac damage and pyroptosis of cardiomyocytes in diabetic mice. LIFE SCIENCES (PubMed: 33508297) [IF=6.1]

Application: WB    Species: mice    Sample: left ventricle tissues

Fig. 6. Knockdown of SGLT1 suppresses inflammatory response and pyroptosis in GVDM mice. A, Protein levels of NLRP3, ASC and caspase-1 (p20) in left ventricle tissues were measured by western blot. B, Quantitative analysis of relative protein levels of NLRP3, ASC and caspase-1. C, Protein levels of full length GSDMD and cleaved GSDMD were measured by western blot. D, Quantitative analysis of relative protein levels of cleaved GSDMD. E, The expression of NLRP3 was detected by immunofluorescence. Scale bar: 50 μm. F, Caspase-1 activity was measured by commercial kit. The relative mRNA levels of IL-1β (G) and IL-18 (H) were detected by qRT-PCR. I, The protein levels of IL-1β, pro-IL-1β and IL-18 were measured by western blot. Quantitative analysis of relative protein levels of IL-1β (J) and IL-18 (K). The release of IL-1β (L) and IL-18 (M) were detected by ELISA. Data were expressed as mean ± Standard deviation (SD). ***p < 0.001, **p < 0.01, *p < 0.05 vs control mice; †††p < 0.001, ††p < 0.01, † p < 0.05 vs DM mice; ###p < 0.001, ##p < 0.01 vs GVDM+shNC mice. DM, diabetes mellitus; GV, glycemic variability; SGLT1, sodium-glucose cotransporter 1; shSGLT1, SGLT1 shRNA; shNC, negative control shRNA; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing CARD domain; GSDMD, gasdermin D.
3). Activation of ATM/Chk2 by Zanthoxylum armatum DC extract induces DNA damage and G1/S phase arrest in BRL 3A cells. JOURNAL OF ETHNOPHARMACOLOGY (PubMed: 34775036) [IF=5.4]
4). Regulation of Signaling Pathways Involved in the Anti-proliferative and Apoptosis-inducing Effects of M22 against Non-small Cell Lung Adenocarcinoma A549 Cells. Scientific Reports (PubMed: 29343765) [IF=4.6]

Application: WB    Species: human    Sample: A549 cells

Figure 3. |M22 regulates G0/G1 arrest through a negative feedback mechanism. (A) M22 attenuates mRNA expression of Cyclin D1, Cyclin E1, CDK4, CDK6, CDC25A and PCNA genes in A549 cells for 24 h. RNA was isolated and analsis was performed to detect by qRT-PCR. *Means P < 0.01, ***means P < 0.0001. (B) M22 down-regulates the expression of Cyclin D1 and CDC25C protein in A549 cells by Western blotting.
5). Overexpression of SMYD3 in Ovarian Cancer is Associated with Ovarian Cancer Proliferation and Apoptosis via Methylating H3K4 and H4K20. Journal of Cancer (PubMed: 31417652) [IF=3.9]

Application: WB    Species: human    Sample: HEY and A2780 cells

Figure 5. |SMYD3 regulated ovarian cancer proliferation and apoptosis though SMYD3-H4K20me3-CDKN2A and SMYD3-H3K4me3-BIRC3, respectively. (A-B) The protein expression changes in the above 6 genes in the HEY and A2780 cell lines after SMYD3 knockdown in HEY and A2780 cells.
6). 6,12-Diphenyl-3, 9-diazatetraasterane-1, 5, 7, 11-tetracarboxylate Inhibits Proliferation, Migration and Promotes Apoptosis in Ovarian Cancer Cells. DISEASE MARKERS (PubMed: 32963636)

Application: WB    Species: human    Sample: SKOV3 cells

Figure 5: |The effect of DDTC on expression of cell cycle regulators. The dosage of DDTC at 0.1 μM, 1 μM, and 10 μM in SKOV3 and A2780 cells. (a, b) Western blot assay showed that DDTC upregulated the expression of Chk1 protein level and downregulated the expression of Cdc25a and CDK1protein levels in SKOV3 cells.

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