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Af1830

Manufactured by R&D Systems
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Sourced in Germany

AF1830 is a recombinant human protein product manufactured by R&D Systems. It is a member of the Fibroblast Growth Factor (FGF) family.

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Lab products found in correlation

Provisc, by Alcon (1 mentions) Ultra fine, by BD (1 mentions) Mab002, by R&D Systems (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Rabbit anti actin, by Merck Group (1 mentions) Rabbit anti gapdh, by Cell Signaling Technology (1 mentions) Ab47027, by Abcam (1 mentions) Mab377, by Merck Group (1 mentions) Mbp84 104, by GenScript (1 mentions) Mab5262, by Merck Group (1 mentions) Sc 188, by Santa Cruz Biotechnology (1 mentions) Af506, by R&D Systems (1 mentions) Il 6r, by R&D Systems (1 mentions) Glial fibrillary acidic protein (gfap), by Agilent Technologies (1 mentions) Ab 108 c, by R&D Systems (1 mentions) Ab133357, by Abcam (1 mentions) Tyr705, by Cell Signaling Technology (1 mentions) Ab15580, by Abcam (1 mentions) Nbp2 12506, by Novus Biologicals (1 mentions) Ab3849, by Merck Group (1 mentions)

5 protocols using af1830

1

Induction of Retinal Detachment for Research

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A previous method [12 ] for inducing bullous and permanent RD was modified to avoid any retinal holes and subretinal hemorrhage. Briefly, a superior scleral hole was gently made with the bevel of 30 G insulin syringe needle (BD Ultra-Fine™), avoiding any retinal damage. Anterior chamber puncture was made to relieve any elevation of intraocular pressure. A 33 G needle connected to Hamilton 2.5 μl syringe was inserted into the scleral hole and carefully positioned within the subretinal space. Sodium hyaluronate (2 μl, ProVisc, Alcon) was gently injected to detach the neurosensory retina from RPE. In the treated groups, 2 μl of polyclonal goat anti-mouse IL6Rα IgG antibody (R&D system, AF1830) or monoclonal rat anti-mouse gp130 IgG1 antibody (R&D system, MAB4682) or monoclonal mouse IgG1 isotype control (R&D system, MAB002) or PBS was injected into the vitreous cavity following detachment. The success of hemi-retinal detachment was confirmed by visual microscopic inspection, and in a subset of cases, OCT imaging. Mice with detachments that were accompanied by retinal holes, subretinal or vitreous hemorrhage were excluded from subsequent studies. More details about anti-IL6Rα and anti-gp130antibodies used in the current study are given in Additional file 1: Table S1.
Wang X., Miller E.B., Goswami M., Zhang P., Ronning K.E., Karlen S.J., Zawadzki R.J., Pugh EN J.r, & Burns M.E. (2017). Rapid monocyte infiltration following retinal detachment is dependent on non-canonical IL6 signaling through gp130. Journal of Neuroinflammation, 14, 121.
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2

Protein Expression and Localization

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Primary antibodies included rabbit-anti-CT-Ab (#SC661; Santa Cruz), rabbit-anti-actin (A2066, Sigma-Aldrich, Steinheim, Germany), rabbit-anti-GAPDH (14C10, Cell Signaling, Frankfurt am Main, Germany), rabbit-anti-meprin α (self-designed at Pineda, Berlin, Germany), and rabbit-anti-meprin β72 (link), mouse-anti-human-IL-6R (4–11)73 (link) and goat-anti-mouse-IL-6R (AF1830, R&D systems, Wiesbaden-Nordenstadt, Germany). Secondary antibodies used were Alexa Fluor 488, biotinylated antibodies (Molecular Probes, Eugene, USA) and peroxidase conjugated antibodies (Dianova, Hamburg, Germany).
Arnold P., Boll I., Rothaug M., Schumacher N., Schmidt F., Wichert R., Schneppenheim J., Lokau J., Pickhinke U., Koudelka T., Tholey A., Rabe B., Scheller J., Lucius R., Garbers C., Rose-John S, & Becker-Pauly C. (2017). Meprin Metalloproteases Generate Biologically Active Soluble Interleukin-6 Receptor to Induce Trans-Signaling. Scientific Reports, 7, 44053.
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3

Immunofluorescence Analysis of Neural Markers

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Routine reagents were purchased from Sigma unless indicated otherwise. MBP84-104 (ENPVVHFFKNIVTPRTPPPSQ) and scrambled (s)MBP84-104 (NKPQTNVVEPFHRTFPIPPVS) peptides, derived from the human MBP sequence (GenBank #AAH08749), were synthesized by GenScript. The peptides were protected from degradation by exoproteinases using N-terminal acetylation and C-terminal amidation. The following primary antibodies were used in our immunofluorescence analyses: goat polyclonal IL-6 [R&D Systems (AF506), 1:100], goat polyclonal IL-6 receptor [IL-6R, R&D Systems (AF1830), 1:100], rabbit polyclonal glial fibrillary acidic protein [GFAP, DAKO (Z0334), 1:500], mouse monoclonal NeuN [EMD Millipore (MAB377), 1:1000], rabbit ionized Ca2+-binding adapter molecule 1 [Iba1, Wako (019-19741), 1:500], mouse neurofilament 200 [NF200, Millipore (MAB5262), 1:200], rabbit polyclonal calcitonin gene-related peptide [CGRP, Abcam (ab47027), 1:400], and rabbit polyclonal activating transcription factor 3 [ATF3, C-19 clone, Santa Cruz Biotechnology (SC-188), 1:100].
Ko J.S., Eddinger K.A., Angert M., Chernov A.V., Dolkas J., Strongin A.Y., Yaksh T.L, & Shubayev V.I. (2016). Spinal activity of interleukin 6 mediates myelin basic protein-induced allodynia. Brain, behavior, and immunity, 56, 378-389.
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4

Macrophage Infection Model with FAC

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pMacs in a 24‐well tissue culture plate (∼5 × 105 cells/well, 1 mL culture media/well) were treated for 24 h with “infection medium,” which was culture medium lacking penicillin/streptomycin, with or without 250 μM ferric ammonium citrate (66 μg/mL, FAC, Sigma–Aldrich, CAS: 1185‐57‐5) and with or without 125 ng/mL anti‐IL‐6Rα (R&D Systems, AF1830) or normal goat IgG isotype control antibody (R&D Systems, AB‐108‐C). Cells were infected with UTI89 in 1 mL of new infection media.
Owusu‐Boaitey N., Bauckman K.A., Zhang T, & Mysorekar I.U. (2016). Macrophagic control of the response to uropathogenic E. coli infection by regulation of iron retention in an IL‐6‐dependent manner. Immunity, Inflammation and Disease, 4(4), 413-426.
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5

Comprehensive Immunohistochemistry Protocol

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Formalin‐fixed, paraffin‐embedded tissues were cut in 5‐μm sections. Sections were evaluated by immunohistochemical analysis following our previously reported protocol (Sorrelle et al, 2019) using antibodies specific for anti‐P‐STAT3 (Tyr705, #9145, Cell Signaling, dilution 1:500), anti‐ARG1 (#93668, Cell Signaling, dilution 1:500), anti‐CD8α (#98941, Cell Signaling, dilution 1:1,000), anti‐ZAP70 (#2705, Cell Signaling, dilution 1:500), anti‐P‐SMAD2 (Ser465/467, AB3849, Millipore, dilution 1:500), anti‐IL‐6Rα (AF1830, R&D Systems, dilution 1:250), anti‐iNOS (PA1‐21054, Thermo Fisher Scientific, dilution 1:200), anti‐CD3 (PA1‐29547, Thermo Fisher Scientific, dilution 1:1,000), anti‐F4/80 (NBP2‐12506, Novus Biologicals, 1:200), anti‐FOXP3 (MAB8214, Novus Biologicals, dilution 1:500), anti‐CD11b (ab133357, Abcam, dilution 1:1,000), and anti‐Ki67 (ab15580, Abcam, dilution 1:1,000). Images were obtained with a Nikon Eclipse E600 microscope using a Niko Digital Dx1200me camera and ACT1 software (Universal Imaging Corporation). Pictures were analyzed using NIS Elements (Nikon).
Huang H., Zhang Y., Gallegos V., Sorrelle N., Zaid M.M., Toombs J., Du W., Wright S., Hagopian M., Wang Z., Hosein A.N., Sathe A.A., Xing C., Koay E.J., Driscoll K.E, & Brekken R.A. (2019). Targeting TGFβR2‐mutant tumors exposes vulnerabilities to stromal TGFβ blockade in pancreatic cancer. EMBO Molecular Medicine, 11(11), e10515.
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