ADAMTS13 Overexpression Lysate

Name: ADAMTS13 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-05570

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-05570

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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 ADAMTS13 Overexpression Lysate

ADAMTS13 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC220320

Accession#: NM_139025

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

149.9 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 ADAMTS13 Overexpression Lysate

Western Blot: ADAMTS13 Overexpression Lysate [NBP2-05570]

Western Blot: ADAMTS13 Overexpression Lysate (Adult Normal) [NBP2-05570] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ADAMTS13.

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: ADAMTS13

ADAMTS13 was first identified by its ability to cleave the ultra large aggregates of vWF into smaller forms in the serum. A thrombocytopenic disorder called thrombic thrombocytopenic purpura (TTR) was shown to be due to a deficiency in the cleavage of von Willebrands factor, and ADAMTS13 was identified as the enzyme responsible. ADAMTS13 cleaves the Tyr1605 to Met1606 bond in the A2 domain of vWF, a process stimulated by shear stress in the circulation and by binding to either platelet glycoprotein IBa or to heparin. The cleaved vWF leads to decreased platelet adhesion. An idiopathic form of TTR is found in patients with auto antibodies to ADAMTS13. The full length ADAMTS-1 sequence codes for a 1,427 amino acid protein, with a predicted mass of 153.6 kDa, but glycosylation and the abundance of cysteine residues gives ADAMTS-13 an apparent molecular weight of about 190 kDa on reduced SDS PAGE gels. The variants reported include 1,371, 1,340 and 344 amino acids, with predicted mass of 130.3, 144.5 and 36.7 kDa respectively. Many bands of varying sizes can be seen on Western blots, between 110-190 kDa, perhaps indicating differentially processed ADAMTS-13.

Long Name

A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 13

Alternate Names

TTP, vWF-CP

Gene Symbol

ADAMTS13

Additional ADAMTS13 Products

Product Documents for ADAMTS13 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for ADAMTS13 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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