Cyclin E1 antibody

Catalog No. ABIN967324

Product Details anti-Cyclin E1 Antibody

Target: Cyclin E1 (CCNE1)

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This Cyclin E1 antibody is un-conjugated

Application: Western Blotting (WB), Flow Cytometry (FACS), BioImaging (BI)

Brand: BD Pharmingen™

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
3. Triton is a trademark of the Dow Chemical Company.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
6. Please refer to us for technical protocols.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Clone: HE12

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968535, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1.0 mg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-Cyclin E1 antibody (ABIN967324)

Darzynkiewicz, Gong, Juan, Ardelt, Traganos: "Cytometry of cyclin proteins." in: Cytometry, Vol. 25, Issue 1, pp. 1-13, (1997) (PubMed).

Target Details for Cyclin E1

Target: Cyclin E1 (CCNE1)

Alternative Name: Cyclin E (CCNE1 Products)

Synonyms: CCNE1 antibody, ccne antibody, cyclinE antibody, CCNE antibody, AW538188 antibody, CycE1 antibody, CYCLE antibody, Ccne antibody, cyce2 antibody, cycE3 antibody, cyce antibody, fa18e07 antibody, wu:fa18e07 antibody, cyclin E1 antibody, cyclin E1 L homeolog antibody, cyclin E1 S homeolog antibody, CCNE1 antibody, ccne1 antibody, Ccne1 antibody, ccne1.L antibody, ccne1.S antibody

Background: Cyclins and cyclin-dependent kinases (cdks) are evolutionarily conserved proteins that are essential for cell-cycle control in eukaryotes. Cyclins (regulatory subunits) bind to cdks (catalytic subunits) to form complexes that regulate the progression of the cell cycle. These complexes are regulated by activating and inhibitory phosphorylation events, as well as by interactions with small proteins that bind to cyclins, cdks, or cyclin-cdk complexes, e.g., p21 and p27 [Kip1]. Cyclin E is expressed in G1 and associates with cdk2 to form an active kinase where it plays an important role in the regulation of the G1/S restriction checkpoint in the cell cycle. Abberant expression of cyclin E has been reported to be associated with the oncogenic transformation of cells. This antibody has been reported not to cross-react with mouse cyclin E.

Molecular Weight: 50 kDa

Pathways: Cell Division Cycle, Intracellular Steroid Hormone Receptor Signaling Pathway, Nuclear Hormone Receptor Binding, Mitotic G1-G1/S Phases