Recombinant Human Interferon gamma protein (Active)

• FuncS

Supplier Data

Functional Studies - Recombinant Human Interferon gamma protein (Active) (AB259377)

Fully biologically active determined by the dose dependent reduction in cytopathic effect of viral infection in A549 cells. ED₅₀ for this effect is 0.30 ng/ml corresponding to a specific activity of 3.33 x 10⁶ units/mg.

• Mass Spec

Lab

Mass Spectrometry - Recombinant Human Interferon gamma protein (Active) (AB259377)

Mass Spectrometry analysis of ab259377 (Agilent Technologies). The mass spectrum shows the full length protein and two C-terminal truncated versions (-5 aa and -16 aa). Heterogeneity of C-terminus has been reported in literature (PAN, Yu‐Ching E., et al. "Structural characterization of human interferon γ Heterogeneity of the carboxyl terminus." European journal of biochemistry 166.1 (1987) : 145-149. ).

• Mass Spec

Supplier Data

Mass Spectrometry - Recombinant Human Interferon gamma protein (Active) (AB259377)

M + 1.4 Da (calc, mass 16833.6)

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).

• sELISA

Lab

Sandwich ELISA - Recombinant Human Interferon gamma protein (Active) (AB259377)

Wild-type A549 control cells or IP-10 knockout A549 cells (ab266969), grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (ab259377) at 100 ng/ml and Recombinant human TNF alpha protein (ab259410) at 10 ng/ml or vehicle control for 16 or 32 hours.

THP-1 cells, grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (ab259377) at 200 ng/ml and LPS at 50 ng/mL or vehicle control for 24 hours.

The concentrations of IP-10 (CXCL10) in cell culture supernatants were measured in duplicate and interpolated from the IP-10 standard curves using Human IP-10 ELISA Kit (ab173194). IP-10 from vehicle control samples were measured in undiluted supernatants and the treated samples were diluted 200 times. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

• HPLC

Supplier Data

HPLC - Recombinant Human Interferon gamma protein (Active) (AB259377)

Purity : 100%

The spectrum was recorded using a 1260 Infinity II HPLC system with DAD (Agilent Technologies) and a MabPac RP column (3.0x100 mm, 4 μm, Thermo Scientific). 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.

• SDS-PAGE

Supplier Data

SDS-PAGE - Recombinant Human Interferon gamma protein (Active) (AB259377)

SDS-PAGE analysis of ab259377.