Recombinant human IL-6 protein (Active)

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Recombinant human IL-6 protein (Active) (AB259381)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6 antibody (ab323372) or isotype IgG (ab210849).​

Cell proliferation (%) (A) confirms that the anti-IL-6 antibody (ab323372) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 4.990 ng/ml (B). ​

TF-1 cells were seeded at 5 × 10⁴ cells per well in a 96-well plate and cultured in medium containing 5% FBS for 48 h. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Recombinant human IL-6 protein (Active) (AB259381)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6 antibody (ab322419) or isotype IgG (ab210849). Cell proliferation (%) (A) confirms that the anti-IL-6 antibody (ab322419) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 1.450 μg/ml (B). ​ TF-1 cells were seeded at 5 × 10⁴ cells per well in a 96-well plate and cultured in medium containing 5% FBS for 48 h. Luminescence was recorded using a Microplate Reader at 560nm emission. ​

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Recombinant human IL-6 protein (Active) (AB259381)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6R antibody (ab323483) or isotype IgG (ab210849).

Cell proliferation (%) (A) confirms that the anti-IL-6R antibody (ab323483) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 0.1914 ug/ml (B).

TF-1 cells were plated at a density of 5 × 10⁴ cells per well in a 96-well plate. A dilution series of the anti-IL-6R antibody (ab323483) was added to the cells and pre-incubated for 2 hours before IL-6 (ab259381) stimulation, and cells were cultured in medium with 5% FBS for a duration of 48 hours. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.

• sELISA

Lab

Sandwich ELISA - Recombinant human IL-6 protein (Active) (AB259381)

Sandwich ELISA - Recombinant human IL-6 protein standard curve.

Background subtracted standard curve using Human IL-6 Antibody Pair - BSA and Azide free (ab243973) and Recombinant human IL-6 protein (Active) (ab259381) 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.

• Mass Spec

Lab

Mass Spectrometry - Recombinant human IL-6 protein (Active) (AB259381)

M+1 Da, pass (+203 Da : Hex, +203 : HexNAc, +291 : NeuAc).

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).

• FuncS

Lab

Functional Studies - Recombinant human IL-6 protein (Active) (AB259381)

Fully biologically active when compared to standard. The ED₅₀ as determined by the dose-dependant Proliferation of TF-1 cells is 0.8067 ng/mL corresponding to a specific activity of 1.24 x 10⁶ IU/mg.

• SDS-PAGE

Lab

SDS-PAGE - Recombinant human IL-6 protein (Active) (AB259381)

SDS-PAGE analysis of ab259381.

• HPLC

Lab

HPLC - Recombinant human IL-6 protein (Active) (AB259381)

Purity 99%.

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.