Detection of Human Cyclin E1 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Cyclin E1 Monoclonal Antibody (Catalog # MAB68101) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Cyclin E1 at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Cyclin E1
Cyclin E1 (also known as G1/S-specific cyclin-E1 and CCNE1) is a 48-58 kDa member of the cyclin E subfamily, cyclin family of molecules. It associates with Cdk2 kinase, and determines substrate specificity for the complex. This complex phosphorylates multiple substrates involved in cell cycle progression and the initiation of DNA replication. Cyclin E1 is required for G1/S phase progression and cell cycle reentry from G0 phase. Its reduced activity during cellular senescence contributes to G1 arrest. Human cyclin E1 is 410 amino acids (aa) in length. It contains two cyclin box folds (aa 144-234 and 293-363) and is phosphorylated on at least eight Ser/Thr sites. There are at least two alternate splice forms. One is 40-41 kDa in size and utilizes an alternate start site at Met46, while a second is 43-44 kDa in size and shows a deletion of aa 154-196. One other potential start site exists at Met16. There are multiple proteolytic cleavage products that are tumor-associated and show increased destabilizing activity. Cleavage around Gln40 generates 44-45 kDa C-terminal fragments, while cleavage between Ala69Asp70 generates 33-35 kDa C-terminal fragments. Over aa 16-143, human cyclin E1 shares 67% aa identity with mouse cyclin E1.
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