Leaf extract (1 mg/ml in PBS), TGFβ1 antibody (5 µg/ml in PBS; clone 1D11 Bio-Techne), parthenium (1 mg/ml in chloroform; Sigma), artemisinin (1 mg/ml in chloroform; Sigma), full length Pisum sativum L. (Leguminosae) chlorophyll a-b binding protein AB96 (at indicated concentrations; mybiosource TTKKVASSSSPWHGPDGVK YLGPFSGESPSYLTGEFPGDY GWDTAGLSADPETFAK NRELEVIHSRWAMLGAL GCVFPELLSRNGVKFG EAVWFKAGSQIFSEGGL DYLGNPSLVHAQSIL AIWATQVILMGAVEGYRI AGGPLGEVVDPLYPG GS FDPLGLAEVPEAFAE LKVKELKNGRLAMF SMF GFFVPAIVTGKGPLEN LADHLADPVNNN AWSYATNFVPGK), soybean trypsin inhibitor (at indicated concentrations; Sigma) or vehicle (PBS or chloroform) were plated on a high binding 96 well flat-bottomed plate (Greiner) and left at RT overnight. The next day the plate was washed × 3 in PBS + 0.05% Tween-20 (Sigma) before being blocked with 5% Tween-20 in PBS. Active TGFβ1 (Bio-Techne), IL-10 (Bio-Techne), IL-12p70 (Bio-Techne) or latent TGFβ1 (Bio-Techne) were added at indicated concentrations before being incubated for 2 h at RT. The supernatant was removed and placed immediately in a relevant ELISA for assaying; this formed the ‘unbound fraction’. The plate was washed × 3 in PBS + 0.05% Tween-20 before being treated with 50 µl 1 M HCl for 30 min at RT to elute any bound protein. This ‘bound fraction’ was neutralised using 50 µl 1.2 M NaOH/0.5 M HEPES and immediately assayed in a relevant ELISA.
Il 12p70
Manufactured by Bio-Techne
Sourced in United Kingdom
IL-12p70 is a recombinant human cytokine. It is a heterodimeric protein composed of p35 and p40 subunits. IL-12p70 is a key regulator of cell-mediated immunity and plays a critical role in the differentiation of T helper 1 (Th1) cells.
Automatically generated - may contain errors
Lab products found in correlation
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M csf, by Bio-Techne (2 mentions)
Pai 1, by Bio-Techne (2 mentions)
Epidermal growth factor (egf), by Bio-Techne (2 mentions)
Phospho shp2, by Bio-Techne (2 mentions)
Tgf β1, by Bio-Techne (2 mentions)
Nanodrop 2000c, by Thermo Fisher Scientific (2 mentions)
Artemisinin, by Merck Group (1 mentions)
Active tgfβ1, by Bio-Techne (1 mentions)
Tween 20, by Merck Group (1 mentions)
Soybean trypsin inhibitor, by Merck Group (1 mentions)
Il 10, by Bio-Techne (1 mentions)
Luminex is 200 instrument, by Bio-Rad (1 mentions)
Il 1β, by Bio-Techne (1 mentions)
Mmp 9, by Bio-Techne (1 mentions)
Tumor necrosis factor (tnf), by Bio-Techne (1 mentions)
4 protocols using il 12p70
1
Quantifying TGFβ1 binding to leaf extracts
2
Cytokine and Growth Factor Assay Protocol
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3
Cytokine and Growth Factor Assay Protocol
4
Multiplexed Cytokine Profiling of Serum and Saliva
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Serum and saliva samples were thawed on ice, vortexed, and spun down at 16,000× g for 5 min at 4 °C. The samples were diluted (serum 1:1 and saliva 2:1) with the RD1-W buffer (R&D Systems, Abington, UK). All samples were analyzed with the custom-made 12-plex Luminex Mouse Discovery Assay kit (http://www.biotechne.com/g8AnddcM (accessed on 31 March 2022)) including CCL3/MIP-1α, KC, IP-10, G-CSF, IFN-γ, IL-1α, IL-1β, IL-6, IL-12 p70, MMP-9, TIMP-1, and TNF (Bio-Techne Ltd., Abington, UK). The plates with saliva samples were incubated overnight. A Luminex IS 200 instrument (Bio-Rad, Hercules, CA, USA) was used to record data. IL-1β, IL-6, IP-10, or IFN-γ were below the level of detection in saliva samples, while MMP-9 was excluded for both serum and saliva samples as the data were above the standard curve. Due to high background levels, IL-12 p70 was also excluded from the cytokine panel. Total protein concentrations in saliva samples were measured in mg/mL using spectrophotometry (Absorbance 280 nm, NanoDrop 2000c, Thermo Fisher Scientific, Waltham, MA, USA). The salivary cytokine levels were adjusted to total protein concentration and presented as (pg of cytokine)/(mg of total protein).
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