Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E303] to Cdc25C
- Suitable for: WB, Flow Cyt, IP
- Knockout validated
- Reacts with: Human
Product nameAnti-Cdc25C antibody [E303]
See all Cdc25C primary antibodies
DescriptionRabbit monoclonal [E303] to Cdc25C
Specificityab32050 recognises Cdc25C. It does not cross react with other MPI phosphatase family members.
Tested applicationsSuitable for: WB, Flow Cyt, IPmore details
Unsuitable for: ICC/IF or IHC
Species reactivityReacts with: Human
Synthetic peptide within Human Cdc25C aa 450-550 (C terminal). The exact sequence is proprietary.
- WB: HeLa, Hap1 and A431 cell lysates.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
KO cell lines
KO cell lysates
Our Abpromise guarantee covers the use of ab32050 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 53 kDa).|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionFunctions as a dosage-dependent inducer in mitotic control. It is a tyrosine protein phosphatase required for progression of the cell cycle. It directly dephosphorylates CDK1 and activate its kinase activity.
Sequence similaritiesBelongs to the MPI phosphatase family.
Contains 1 rhodanese domain.
Developmental stageExpressed predominantly in G2 phase.
modificationsPhosphorylated by CHK1 on Ser-216. This phosphorylation creates a binding site for 14-3-3 protein and inhibits the phosphatase. Phosphorylated by PLK4.
- Information by UniProt
- CDC 25 antibody
- Cdc 25C antibody
- CDC25 antibody
All lanes : Anti-Cdc25C antibody [E303] (ab32050) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CDC25C knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 53 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Lanes 1- 2: Merged signal (red and green). Green - ab32050 observed at 58 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32050 was shown to react with Cdc25C in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265189 (knockout cell lysate ab257387) was used. Wild-type HeLa and CDC25C knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Cdc25C knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human bladder lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32050 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32050 was shown to specifically react with Cdc25C when Cdc25C knockout samples were used. Wild-type and Cdc25C knockout samples were subjected to SDS-PAGE. ab32050 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Anti-Cdc25C antibody [E303] (ab32050) at 1/1000 dilution + A431 cell lysate
Predicted band size: 53 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32050 has been referenced in 3 publications.
- Han L et al. Long non-coding RNA SOX2OT promotes cell proliferation and motility in human ovarian cancer. Exp Ther Med 15:2182-2188 (2018). PubMed: 29434823
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Subramaniam D et al. Translation inhibition during cell cycle arrest and apoptosis: Mcl-1 is a novel target for RNA binding protein CUGBP2. Am J Physiol Gastrointest Liver Physiol 294:G1025-32 (2008). PubMed: 18292181