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VHL (D2N9L) Rabbit Monoclonal Antibody (trFluor Europium Cryptate Conjugate) #24302

    Product Specifications

    REACTIVITY
    SENSITIVITY Endogenous
    MW (kDa)
    Source/Isotype Rabbit IgG

    Product Information

    Product Description

    This Cell Signaling Technology® antibody is conjugated to trFluor™ Europium Cryptate and is expected to detect endogenous human VHL protein. Time-resolved (TR) fluorescence resonance energy transfer (FRET), or TR-FRET, data was generated using the conjugated antibody.

    Fluorescent Properties

    • ← Excitation: 305 nm ← Emission: 617 nm

    Product Usage Information

    trFluor™ Europium Cryptate conjugated antibodies are ideal for protein-protein interaction assays, pair-based immunoassay technologies, or high-throughput screening where greater sensitivity and quantitative detection is needed. The long fluorescent decay time of Europium Cryptate results in improved signal to background over fluorescent dye conjugates. Methods utilizing Europium Cryptate conjugated antibodies include, but are not limited to, TR-FRET, where Europium Cryptate-labeled donor antibodies are paired with red fluorophore-labeled acceptor antibodies, such as Alexa Fluor® 647 or APC. Please contact us if you require an acceptor antibody clone conjugated to a red dye at a custom concentration, a carrier-free formulation, or a more customized packaging solution.

    Note: Addition of potassium fluoride (KF) in the assay dilution buffer range of 40-400 mM is recommended for enhanced fluorescent signal. Optimal dilutions and working concentrations of both the conjugated antibodies and the KF should be determined by the end user.

    Storage

    Supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

    Specificity / Sensitivity

    VHL (D2N9L) Rabbit mAb (trFluor™ Europium Cryptate Conjugate) recognizes endogenous levels of total VHL protein.

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human VHL protein.

    Background

    The von Hippel-Lindau (VHL) protein is a substrate recognition component of an E3 ubiquitin ligase complex containing elongin BC (TCEB1 and TCEB2), cullin 1 (CUL1), and RING-box protein 1 (RBX1) (1-3). VHL protein has been shown to exist as three distinct isoforms resulting from alternatively spliced transcript variants (4). Loss of VHL protein function results in a dominantly inherited familial cancer syndrome that manifests as angiomas of the retina, hemangioblastomas of the central nervous system, renal clear cell carcinomas, and pheochromocytomas (4). Under normoxic conditions, VHL directs the ubiquitylation and subsequent proteasomal degradation of the hypoxia-inducible factor 1α (HIF-1α), maintaining very low levels of HIF-1α in the cell. Cellular exposure to hypoxic conditions, or loss of VHL protein function, results in increased HIF-1α protein levels and increased expression of HIF-induced gene products, many of which are angiogenesis factors such as vascular endothelial growth factor (VEGF). Thus, loss of VHL protein function is believed to contribute to the formation of highly vascular neoplasias (4). In addition to HIF-1α, VHL is known to regulate the ubiquitylation of several other proteins, including tat-binding protein-1 (TBP-1), the atypical protein kinase C (aPKC) lambda, and two subunits of the multiprotein RNA polymerase II complex (RPB1 and RPB7) (5-8). Interactions with elongin BC, RPB1, RPB7, and the pVHL-associated KRAB-A domain-containing protein (VHLaK) suggest that VHL may also play a more direct role in transcriptional repression.

    Alternate Names

    elongin binding protein; HRCA1; Protein G7; pVHL; RCA1; VHL; VHL1; von Hippel-Lindau disease tumor suppressor; von Hippel-Lindau tumor suppressor; von Hippel-Lindau tumor suppressor, E3 ubiquitin protein ligase

    For Research Use Only. Not for Use in Diagnostic Procedures.
    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    trFluor is a trademark of AAT Bioquest, Inc.
    trFluor is manufactured by AAT Bioquest, Inc.
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