Smad5 Overexpression Lysate

Name: Smad5 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-09079

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-09079

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Product Datasheet / COA / SDS

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Smad5 Overexpression Lysate

Smad5 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC224544

Accession#: NM_001001420

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

52.1 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 Smad5 Overexpression Lysate

Western Blot: Smad5 Overexpression Lysate [NBP2-09079]

Western Blot: SMAD5 Overexpression Lysate (Adult Normal) [NBP2-09079] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SMAD5.

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: Smad5

SMADs are members of the MAD-related family of molecules. MAD-related proteins are a family of intracellular proteins that are essential components in the signaling pathways of the serine/threonine kinase receptors of the transforming growth factor beta superfamily (1). SMADs can be divided into receptor-regulated SMADs (R-SMADs: SMAD1, SMAD2, SMAD5, SMAD8 and SMAD9), common-mediator SMAD (co-SMAD: SMAD4), and inhibitory SMADs (I-SMADs: SMAD6 and SMAD7). SMAD1, SMAD5, SMAD8 and SMAD9 have high degrees of homology and antibodies are available that recognize sequences common to all of them. SMAD8 and SMAD9 are typically used as alternate names for one another in the literature. Human SMAD1 is a 465 amino acid protein; GenBank Accession No. AAP36050.1. Human SMAD2 is a 467 amino acid protein; GenBank Accession No. AAC51918.1 Human SMAD3 is a 425 amino acid protein; GenBank Accession No. NP_005893.1 Human SMAD4 is a 552 amino acid protein GenBank Accession No. NP_005350.1. Human SMAD5 is a 465 amino acid protein; GenBank Accession No. AAC50791.1. Human SMAD6 is a 496 amino acid protein; GenBank Accession No. AAH12986.1 Human SMAD7 is a 426 amino acid protein; GenBank Accession No. AAB81354.1 Mouse SMAD8 is a 430 amino acid protein; GenBank Accession No. AAN85445.1 Human SMAD9 is a 430 amino acid protein; GenBank Accession No. NP_005896.1.

Long Name

Mothers Against DPP Homolog 5

Alternate Names

Dwfc, JV5-1, MADH5

Gene Symbol

SMAD5

Additional Smad5 Products

Product Documents for Smad5 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Smad5 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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