Q: I'd like to check toxicity of following products we recently ordered. 1)NBL1-10600 FASLG Overexpression Lysate, NBL1-11652 HNRNPU Overexpression Lysate 2)NB100-1935 CNPase Antibody. Do these products come with SDS buffer containing 2-mercaptoethanol?

A: Both the FASLG Overexpression Lysate (NBL1-10600) and the HNRNPU Overexpression Lysate (NBL1-11652) contain: 1 vial of 100 ug gene specific transient over-expression cell lysate in RIPA buffer and 1 vial of 0.5 ml 2xSDS Sample Buffer (4%SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 5% b-mercaptoethanol).

Fas Ligand/TNFSF6 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-10600

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

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Fas Ligand/TNFSF6 Overexpression Lysate

Fas Ligand/TNFSF6 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC204894

Accession#: NM_000639

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.

Theoretical MW

31.3 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 Fas Ligand/TNFSF6 Overexpression Lysate

Western Blot: Fas Ligand/TNFSF6 Overexpression Lysate [NBL1-10600]

Western Blot: Fas Ligand Overexpression Lysate (Adult Normal) [NBL1-10600] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Fas Ligand.

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: Fas Ligand/TNFSF6

Fas / APO-1 (CD95) is an important member of the tumour necrosis factor (TNF) superfamily involved in membrane-mediated apoptosis. Ligation of Fas by Fas Ligand (Fas-L) or an anti-Fas cross-linking antibody, triggers activation of the caspase cascade. Functional impairment of the Fas / Fas-L system is associated with the development and progression of malignancies. Fas gene mutations have been suggested to have a role in testicular germ cell tumours. Tumour cells frequently exhibit de novo expression of Fas Ligand (Fas-L), which plays a significant role in local tissue destruction, metastatic spread, and immune escape of the tumor cells. The apoptosis of lymphocytes, which occurs in autoimmune diseases, is usually induced by the Fas/Fas-L system. Fas is believed to be involved in various autoimmune diseases including, ulcerative colitis, Graves disease, and rheumatoid arthritis. Fas expression on gastric epithelial cells in patients infected with H.Pylori is responsible for the accelerated apoptosis of the cells. Serum Fas-L concentration has also been shown to be associated with atherosclerosis and inflammatory disease, in patients with hypertension.

Alternate Names

CD178, CD95L, FASLG, TNFSF6

Gene Symbol

FASLG

Additional Fas Ligand/TNFSF6 Products

Product Documents for Fas Ligand/TNFSF6 Overexpression Lysate

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Certificate of Analysis

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Product Specific Notices for Fas Ligand/TNFSF6 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.

Related Research Areas

Epithelial Cell Markers and Intracellular Molecules

gamma delta T Cells

Isolation, Culture, and Identification of Natural Killer Cells

Natural Killer T (NKT) Cells

Regulation of Apoptosis by TNF Superfamily Members

TNF Superfamily

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