For Il6 neutralizing experiments Il6 neutralizing antibody (R&D, AB-406-NA) and control IgG antibody (R&D, AB-108) were used to assess effects of secreted Il6 on the proliferation of MEFs. The Stat3 inhibitor WP1066 (Selleckchem, S2796), PI3K inhibitor LY294002 (Cell Signaling, #9901), Rapamycin (Cell Signaling, #9904) and JAK inhibitor I (Calbiochem, 420099) were resuspended in DMSO. JQ1 was a kind gift from Dr. J. Bradner (Harvard Medical School) and was used as described previously (31 (link)). Note that the IC50 value for inhibition of proliferation was 1.4 μM for Pten/Trp53-deleted cells, and 3.3 μM for wt cells. These results are in similar range to earlier reported, WP1066 effects on cell survival, proliferation and Stat3 phosphorylation (54 (link)–56 (link)).
Ab 406 na
Manufactured by R&D Systems
Sourced in United States
AB-406-NA is a laboratory equipment product from R&D Systems. It serves a core function in the research and analysis processes, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ly294002, by Cell Signaling Technology (1 mentions)
Rapamycin, by Cell Signaling Technology (1 mentions)
Ab 108, by R&D Systems (1 mentions)
Wp1066, by Selleck Chemicals (1 mentions)
Ab 100 na, by R&D Systems (1 mentions)
Myog 67082 1 ig, by Proteintech (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Dcfh da, by Merck Group (1 mentions)
3 3 diaminobenzidine substrate, by Vector Laboratories (1 mentions)
Vectastain abc peroxidase system, by Vector Laboratories (1 mentions)
Imagescope software, by Leica Biosystems (1 mentions)
Scanscope cs slide scanner, by Leica Biosystems (1 mentions)
Rabbit igg, by Southern Biotech (1 mentions)
5 protocols using ab 406 na
1
Proliferation Effects of IL6 Neutralization
2
Neutralizing TGF-β and IL-6 in Mice
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To neutralize TGF-β, mice were treated with a single dose of a polyclonal TGF-β-neutralizing antibody (AB-100-NA; 100 μg/mouse in 100 μl saline; R & D Systems) at 2 d.p.i. by intraperitoneal (i.p.) injection (7 (link)). To neutralize IL-6, mice were treated i.p. with a single dose of a polyclonal antibody to IL-6 (AB-406-NA; 0.5 μg/mouse in 100 μl saline; R & D Systems) at 4 d.p.i. (56 (link)). Controls were treated i.p. with rabbit IgG at both time points (100 μg/mouse in 100 μl saline; Southern Biotech, Birmingham, AL).
3
Antibody-Mediated Inhibition Assay
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Primary Abs against MyoD (18943-1-AP) and MyoG (67082-1-Ig) were purchased from Proteintech (Wuhan, Hubei, China). The monoclonal neutralization Abs against IL-6 (AB-406-NA) and TNFα (AB-410-NA), and recombinant mouse proteins IL-6 and TNFα, were purchased from R&D Systems (Minneapolis, MN, USA). Fluorescein (FITC)-conjugated AffiniPure goat anti-mouse IgG (H+L) was obtained from KeyGEN Biotechnology (Nanjing, Jiangsu, China). DCFH-DA, NAC, tBHP, DPI and ROT were purchased from Sigma-Aldrich.
4
Immunohistochemical Detection of IL-6 in Lung Tissue
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Thin sections (3 μm) were prepared from formalin-fixed, paraffin-embedded lung tissue (31 (link)). IL-6 was detected using a goat polyclonal antibody (AB-406-NA; R & D Systems, Minneapolis, MN). Bound antibody was detected using biotinylated anti-goat immunoglobulin (Vector Laboratories, Burlingame, CA), the Vectastain ABC peroxidase system, and 3,3′-diaminobenzidine substrate (Vector Laboratories). Sections were counterstained with Harris hematoxylin, scanned with a Scanscope CS slide scanner (Aperio Technologies, Vista, CA), and visualized with ImageScope software (Aperio Technologies).
5
Modulating Immune Responses in Melanoma
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To deplete NK cells, mice received anti-Asialo GM 1 antibodies (1 mg/ml, EBio-Science) or isotype antibodies as control (Rabbit IgG, 0.5 mg/ml, Southern Biotech) every second day from 1 week before B16 inoculation. Five days after tumor cell inoculation, NK cells content was evaluated. Propranolol/Epinephrine Study Eight-week-old C57BL/6 mice were randomized to drinking water containing 0.5 g/l propranolol or nothing. Propranolol administration started 1 week before B16 inoculation. An additional group received daily (Monday to Friday) injections of epinephrine (0.5 mg/kg, 200 ml i.p.) from 1 week before B16 inoculation. For the acute effect of epinephrine, mice were injected with 0.5 mg/kg or 2 mg/kg epinephrine, and sacrified after 30 min, where blood, spleen, and muscle tissue were collected. Anti-IL-6/IL-6 Study Eight-week-old C57BL/6 mice were randomized to groups, receiving i.p. injections of anti-IL-6 antibodies (100 mg/mouse, R&D systems, #AB-406-NA) or vehicle injections twice a week from 1 week before B16 inoculation. An additional group received daily (Monday to Friday) injections of IL-6 (100 ng/mouse, R&D systems, #406-ML-025/CF) from 1 week before B16 inoculation.
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