PRODUCT CODE: ET1612-31

Phospho-Cyclin E1 (T77) Recombinant Rabbit Monoclonal Antibody [SD2025] (ET1612-31)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

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ICC staining of Phospho-Cyclin E1(T77) in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Cyclin E1(T77) in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Cyclin E1(T77) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Cyclin E1(T77) in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-Cyclin E1(T77) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-31, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Phospho-Cyclin E1(T77) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-31, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC staining of Phospho-Cyclin E1(T77) in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-Cyclin E1 (T77) Recombinant Rabbit Monoclonal Antibody [SD2025] (ET1612-31)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding thr77 of human cyclin e1.

Host

Rabbit

Modification

Phospho

Modification Site

T77

Positive Control

SW480, Hela, HepG2, human tonsil tissue, human colon carcinoma tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD2025

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

48 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-Cyclin E1 (T77)

SYNONYMS

CCNE antibody; Ccne1 antibody; CCNE1_HUMAN antibody; cyclin E variant ex5del antibody; cyclin E variant ex7del antibody; Cyclin E1 antibody; Cyclin Es antibody; Cyclin Et antibody; CyclinE antibody; G1/S specific cyclin E antibody; G1/S-specific cyclin-E1 antibody

SEQUENCE SIMILARITIES

Belongs to the cyclin family. Cyclin E subfamily.

TISSUE SPECIFICITY

Highly expressed in testis and placenta. Low levels in bronchial epithelial cells.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation of both Thr-395 by GSK3 and Ser-399 by CDK2 creates a high affinity degron recognized by FBXW7, and accelerates degradation via the ubiquitin proteasome pathway. Phosphorylation at Thr-77 creates a low affinity degron also recognized by FBXW7.; Ubiquitinated by UHRF2; appears to occur independently of phosphorylation.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

Cyclins were first identified in invertebrates as proteins that oscillate dramatically through the cell cycle. These proteins have been well conserved through evolution and play a critical role in regulation of cell division. cyclin E, along with the three cyclin D proteins and cyclin C, has been shown to represent a putative G1 cyclin on the basis of its cyclic pattern of mRNA expression, with maximal levels being detected near the G1/S boundary. cyclin E has been found to be associated with the transcription factor E2F in a temporally regulated manner. The cyclin E/E2F complex is detected primarily during the G1 phase of the cell cycle and decreases as cells enter S phase. E2F is known to be a critical transcription factor for expression of several S phase specific proteins.