Invitrogen™ Cyclin E Monoclonal Antibody (HE12), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-112-4518	25 μg

Catalog No. 50-112-4518 Supplier Invitrogen™ Supplier No. 50971480

Description

Mouse Monoclonal Antibody

The monoclonal antibody HE12 recognizes human Cyclin E. Cyclins regulate the cell cycle by binding and activating cyclin-dependent kinases (CDK). Cyclin E regulates the transition from G1 to S phase by binding and activating CDK2. The Cyclin E/CDK2 complex is responsible for the phosphorylation of downstream proteins that facilitate the G1 to S phase transition. Overexpression of Cyclin E is found in several types of cancer including breast, colon, bladder, lung and skin. In breast cancer, Cyclin E overexpression is associated with increased stage and grade as well as poor prognosis. Applications Reported: This HE12 antibody has been reported for use in immunohistochemical staining of formalin-fixed paraffin embedded tissue sections and microscopy. Applications Tested: This HE12 antibody has been tested by immunohistochemistry of FFPE human tissue using low pH antigen retreival and can be used at less than or equal to 10 μg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor 660 is a replacement for Alexa Fluor 647. eFluor 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

Cyclin E belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. Cyclin E forms a complex with and functions as a regulatory subunit of CDK2, whose activity is required for cell cycle G1/S transition. This protein accumulates at the G1-S phase boundary and is degraded as cells progress through S phase. Overexpression of this gene has been observed in many tumors, which results in chromosome instability, and thus may contribute to tumorigenesis. This protein was found to associate with, and be involved in, the phosphorylation of NPAT protein (nuclear protein mapped to the ATM locus), which participates in cell-cycle regulated histone gene expression and plays a critical role in promoting cell-cycle progression in the absence of pRB. Two alternatively spliced transcript variants of this gene, which encode distinct isoforms, have been described. Two additional splice variants were reported but detailed nucleotide sequence information is not yet available.

Specifications

• Antigen: Cyclin E
• Applications: Immunohistochemistry (Paraffin), Immunocytochemistry
• Classification: Monoclonal
• Clone: HE12
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: CCNE1
• Gene Accession No.: P24864
• Gene Alias: AW538188; Ccne; Ccne1; CycE1; CYCLE; cyclin E; cyclin E1; cyclin Es; cyclin Et; E-S; G1/S-specific cyclin-E1; pCCNE1
• Gene Symbols: CCNE1
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 25 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 898
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov