Primary antibodies: Phospho-STAT3 (Tyr705) (Thermo Fisher, USA; Cat# 44380G), p-NFκB p65 (S536) (Thermo Fisher, USA; Cat# MA515160), STAT3 (Thermo Fisher, USA; Cat# 10253-2-AP), beta Crystallin A3 (Abcam, USA; Cat# ab151722), IL-6 (Biorbyt, USA; Cat# orb6210), IL-1α (Biorbyt, USA; Cat# orb184287) and mNeonGreen (Chromotek, USA; Cat# 3216-100), Secondary antibodies: HRP anti-Rabbit IgG (KPL, USA; Cat# 074-1506), HRP anti-tagged anti-Mouse IgG (KPL, USA; Cat# 5220-0341), HRP anti-tagged Goat IgG (KPL, USA; Cat# 14-13-06). GIPZ Cryba1 shRNA Viral Particle Starter Kit (Dharmacon, USA; Cat# VGH5526-EG12957), AAV2-GFP-U6-shRNA (Vector Biolabs, USA; Cat# 7041). PTP1B (Ad-CMV-mNeonGreen-mPtpn1; Cat# AAV-269791) and Cryba1 (Ad-CMV-RFP-mCryba1; Cat# 2001) overexpression vectors were purchased from Vector Biolabs, USA. ELISA kits: Human crystallin, beta A1 (CRYBA1) (Cat# MBS7252187) and protein tyrosine phosphatase 1B (PTP1B) (Cat# MBS761801) ELISA kits were purchased from Mybiosource, USA. Proteins: CRYBA1 (NM_005208) purified human protein (OriGene Technologies, Cat# TP321965), Recombinant human PTP1B protein (Abcam, Cat# ab51277).
Interleukin 6 (il 6)
Manufactured by Biorbyt
Sourced in United Kingdom
Interleukin-6 (IL-6) is a cytokine involved in immune response and inflammation. It plays a role in the differentiation of B cells and the activation of T cells. IL-6 is a key regulator of the acute phase response.
Automatically generated - may contain errors
Lab products found in correlation
Stat3, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Il 1β, by Biorbyt (1 mentions)
Tnf α, by Biorbyt (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Thincert cell culture insert, by Greiner (1 mentions)
Rankl, by Enzo Life Sciences (1 mentions)
Vascular endothelial growth factor (vegf), by Biorbyt (1 mentions)
Gapdh, by Biorbyt (1 mentions)
Gapdh, by Bio-Rad (1 mentions)
P akt, by Biorbyt (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Imagej software, by Bio-Rad (1 mentions)
4 protocols using interleukin 6 (il 6)
1
Phosphorylation-mediated signaling analysis
2
HNSCC-PBMC Coculture Cytokine Analysis
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HNSCC cells (2 × 105) were seeded in 24-well plates overnight. The next day, PBMCs (3 × 106) were cocultured with the HNSCC cells with/without 10 µg/mL CDA for 2 and 6 hours. For transwell cocultures, 2 × 105 HNSCC cells were seeded into the bottom compartment of a 24-well plate overnight. PBMCs (3 × 106) were seeded into the top 0.4-µm ThinCert cell culture insert (Greiner Bio-One, catalog no. 665641) the next day and cultured for 6 hours ± 10 µg/mL CDA. Supernatants were harvested and the levels of IFNβ, IL6, TNFα, and IL1β were assessed with human type-I IFN (Biorbyt, catalog no. orb561974), TNFα (Invitrogen, catalog no. 88–7346), IL6 (Invitrogen, catalog no. 88–7066), and IL1β (Invitrogen, catalog no. 88–7261) ELISA kits following the manufacturer's instructions.
3
Cytokine Secretion in Prostate Cancer Cells
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DU145 and PC3 cells were seeded onto 24-well plates at a cell count of 2 × 104 cells/well in a regular culture medium and incubated at 37°C for 48 h in the absence and the presence of KPT-330 and KPT- 251 at concentrations as indicated. The conditioned medium was collected and assayed with a sandwich ELISA for TGF-β1 (RayBiotech, Inc., Norcross GA), IL-8 (Biorbyt ltd, Cambridge, UK), IL6 (Biorbyt ltd), VEGF (Biorbyt ltd) and RANKL (Enzo Life Sciences, Inc. Italian distributor Vinci-Biochem Florence) according to the manufacturer’s protocol.
4
Astrocyte Protein Analysis via Western Blot
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Proteins from astrocytes were obtained using the lysis solution (Biorbyt, Cambridge, UK) . The BCA kit (Biorbyt, Cambridge, UK) was used to quantify the isolated proteins, which were loaded on a gel, separated by 12％ SDS-PAGE, and transferred onto the PVDF membrane (Merck, New Jersey, USA) . The membrane was further incubated in 5％ skim milk and incubated at 4℃ overnight with primary antibodies against p-JAK2 (1:800, Biorbyt, Cambridge, UK) , p-STAT3 (1:800, Biorbyt, Cambridge, UK) , p-AKT (1:800, Biorbyt, Cambridge, UK) , GP130 (1:800, Biorbyt, Cambridge, UK) , IL-6 (1:800, Biorbyt, Cambridge, UK) , or GAPDH (1:800, Biorbyt, Cambridge, UK) . Then, the membrane was incubated with secondary antibody (1:4000, Biorbyt, Cambridge, UK) for 2 h. Lastly, following exposure, bands were analyzed using the Image J software (Bio-Rad, California, USA) with GAPDH as the loading control.
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