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SPRED1 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-05326

Novus Biologicals, part of Bio-Techne
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NBP2-05326

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for SPRED1 Overexpression Lysate

SPRED1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC220714

Accession#: NM_152594

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

50.3 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 SPRED1 Overexpression Lysate

Western Blot: SPRED1 Overexpression Lysate [NBP2-05326]

Western Blot: SPRED1 Overexpression Lysate [NBP2-05326]

Western Blot: Spred-1 Overexpression Lysate (Adult Normal) [NBP2-05326] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Spred-1.

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: SPRED1

SPRED1 (Sprouty-related protein with an EVH1 domain-1) is a 55 kDa member of the SPRED family of regulatory molecules. It inhibits mitogenic signaling by blocking Ras activation of Raf, and disrupts actin stress fiber formation by binding TESK1. Human SPRED1 is 444 amino acids (aa) in length. It contains an N-terminal WH1/EVH1 domain that is involved in ERK inhibition (aa 6 - 123), a central KBD domain that binds to the SCFR (aa 233 - 285), and a C-terminal Cys-rich/Sprouty-related domain that mediates homo- and heterodimerization with SPRED2, binding to TESK1, and undergoes palmitoylation for membrane localization (aa 334 - 442). There is one splice variant that shows a nine Lys substitution for aa 269 - 444.

Long Name

Sprouty-related with an EVH1 Domain Containing-1

Alternate Names

NFLS

Gene Symbol

SPRED1

Additional SPRED1 Products

Product Documents for SPRED1 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 SPRED1 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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