MegaTNF-α™ (soluble) (human), (recombinant)



ALX-522-046-C010	10 µg	575.00 USD

Product Specification

Alternative Name:	TNF-α:ACRP30headless, TNFSF 2:ACRP30headless, Tumor necrosis factor α:ACRP30headless

Concentration:	0.1mg/ml after reconstitution.

Endotoxin Content:	<0.1EU/µg purified protein (LAL test; Bio Whittaker).

MW:	~34kDa (SDS-PAGE).

Purity:	≥95% (SDS-PAGE)

Formulation:	Lyophilized. Contains PBS.

Biological Activity:	Killing.

Source/Host:	Produced in HEK 293 cells. The extracellular domain of human TNF-α (tumor necrosis factor-α) (aa 85-233) is fused at the N-terminus to human ACRP30headless (aa 16-108) and a FLAG^®-tag.

Reconstitution:	Reconstitute with 100µl sterile distilled water. Further dilutions should be made with medium containing 5% fetal calf serum.

Specificity:	Binds to human and mouse TNF-R1 and TNF-R2.

Long Term Storage:	-20°C

Use/Stability:	Stable for at least 6 months after receipt when stored at -20°C.

Handling:	After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.

Background / Technical Information:	FLAG is a registered trademark of Sigma-Aldrich Co.

Killing assay using rhsMegaTNF-a^TM on mouse WEHI-164 cells.Method: Murine WEHI 164 cells (50’000 cells in 100µl RPMI medium containing 10% fetal calf serum) were incubated with the indicated concentrations of rhsMegaTNF-a (Prod. No. ALX-522-046) for 12 hours at 37°C in a 96-well plate. Concentrations of rhsMegaTNF-a required to kill cells may vary depending on cell type studied. Cell viability was determined using a PMS/MTS-based cell proliferation assay kit.

rhsMegaTNF-a™ binds to TNF-R1 and TNF-R2 in an ELISA. Binding of rhsMegaTNF-a™ to rhTNF-R1:Fc and rhTNF-R2:Fc as determined by ELISA. Binding to TNF-R1 (and TNF-R2) triggers various biological effects (including cell death). Method: A 96-wells ELISA plate was coated o/n at RT with 100µl of 1µg/ml hTNF-R1:Fc (Prod. No. ALX-522-013) or hTNF-R2:Fc (Prod. No. ALX-522-014) in PBS. 100µl of rhsMegaTNF-a™ at 8µg/ml were mixed with 100µl 0.5% FCS in PBS in the first well and a serial of 1:1 dilution was prepared for the following wells. Bound rhsMegaTNF-a™ to rhTNF-R1:Fc or rhTNF-R2:Fc was revealed using a mouse anti-FLAG antibody (Enhancer, Prod. No. ALX-804-034). ED₅₀ is approx. 0.1ng/ml. The binding of rhsMegaTNF-a™ was compared to rhsTNF-a (Prod. No. ALX-522-008) and 0.5% FCS in PBS was used as a negative control.

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