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NLE1 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13664

Novus Biologicals, part of Bio-Techne
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NBL1-13664

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for NLE1 Overexpression Lysate

NLE1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC204114

Accession#: NM_018096

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

53.1 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for NLE1 Overexpression Lysate

Western Blot: NLE1 Overexpression Lysate [NBL1-13664]

Western Blot: NLE1 Overexpression Lysate [NBL1-13664]

Western Blot: NLE1 Overexpression Lysate (Adult Normal) [NBL1-13664] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NLE1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: NLE1

The Notch signaling pathway is an evolutionary conserved system that is involved in intracellular communication. Notch receptors play an important role in development and cell-fate decisions. Notchless is a loss-of-function mutant allele that encodes for protein NLE1 (notchless homolog 1). NLE1 is a 485 amino acid WD40-repeat protein that binds to the cytoplasmic domain of Notch, regulating its signaling activity in Drosophila melanogaster and in mice. Deletion of the NLE1 gene in mice during the early stages of development results in embryonic death, while gene deletion in the late stages of development leads to activation of a caspase-3-dependent apoptotic pathway.In plants, NLE1 is crucial for normal cellular growth and development. Underexpression during shoot proliferation causes pleiotropic defects such as delayed flowering and abnormal organ maturation. It may also play a role in 60S ribosomal subunit biogenesis in yeast. NLE1 contains eight WD40 domains and produces one isoform due to alternative splicing.

Alternate Names

FLJ10458, Nle, Notchless gene homolog, notchless homolog 1 (Drosophila), notchless protein homolog 1

Gene Symbol

NLE1

Additional NLE1 Products

Product Documents for NLE1 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for NLE1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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