Recombinant mouse TNF alpha protein (Active) (ab259411)

Fully active compared to a standard. The ED₅₀ as determined by the dose-dependant Killing/apoptosis of L-929 cells is 1.12ng/mL corresponding to a Specific Activity of 8.93 x 10⁵ IU/mg.

SDS-PAGE analysis of ab259411.

Purity 100%.

The spectrum was recorded using a 1260 Infinity II HPLC system with DAD and a MabPac RP column (3.0x100 mm, 4 µm). 5 µL of purified protein was injected and the gradient run from 80 % water:TFA (99.9:0.1 v/v) and 20 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) to 20 % water:TFA (99.9:0.1 v/v) and 80 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 3 min. Flow rate was 0.5 mL/min and the column compartment temperature was 50 °C.

M + 1.5 Da (Calc mass 19797.5 Da).

The spectrum was recorded with a 6545XT AdvanceBio LC/Q-TOF (Agilent Technologies) and a MabPac RP column (42.1x50 mm, 4 µm, Thermo Scientific). 5 µL of purified protein was injected and the gradient run from 85 % water:FA (99.9:0.1 v/v) and 15 % acetonitrile:FA (90:9.9:0.1 v/v/v) to 55 % water:FA (99.9:0.1 v/v) and 45 % acetonitrile:FA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 2.5 min. Flow rate was 0.4 mL/min and the column compartment temperature was 60 °C. Data was analysed and deconvoluted using the Bioconfirm software (Agilent Technologies).

Background subtracted standard curve using Mouse TNF alpha Antibody Pair - BSA and Azide free (ab241672) and Recombinant mouse TNF alpha protein (Active) (ab259411) in sandwich ELISA.
The ELISA was performed using the components of the corresponding SimpleStep® kit, which uses the same antibody pair with a different formulation and format.