Fibulin 1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

FIBULIN-1 is a protein that is a component of the extracellular matrix. It plays a role in the structure and function of the basement membrane and elastic fibers.

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Lab products found in correlation

Versican, by Abcam (1 mentions) Normal goat serum (ngs), by Merck Group (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Triton x 100, by Merck Group (1 mentions) Sephacryl s 300, by GE Healthcare (1 mentions) Deae sephadex, by GE Healthcare (1 mentions)

2 protocols using fibulin 1

Immunohistochemical Analysis of Retinal Apoptosis and Oxidative Stress

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Enucleated eyes were fixed in 4% PFA/PBS for 45 min, followed by cryoprotection in 30% sucrose/PBS. Ten μm-embedded frozen sections were further processed for immunohistochemistry. Briefly, frozen retina sections were blocked in PBS with 3% normal goat serum (Sigma, Buchs, Switzerland) and 0.2% Triton X-100 (Sigma) for 1 h at RT and then incubated with antibodies against cleaved Caspase3 (dilution 1/500, Cell Signaling Technology, Inc. Danvers, MA, USA), GPX3 (dilution 1/50, Abcam, Cambridge, MA, USA), GSTO-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA), FIBULIN-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA) and VERSICAN (dilution 1/100, Abcam, Cambridge, MA, USA) in the blocking buffer overnight at 4°C. Thereafter, sections were incubated again in blocking buffer for 30 min at RT before being incubated for 1 h at RT with FITC Alexa-Fluor 594 goat anti-rabbit antibody or with FITC Alexa-Fluor 594 sheep anti-goat antibody (dilution 1/2’000,) depending of primary antibody used. Incubation with secondary antibody alone was used as a negative control. Tissue sections were counterstained with DAPI to identify retinal cell layers.

Emery M., Nanchen N., Preitner F., Ibberson M, & Roduit R. (2016). Biological Characterization of Gene Response to Insulin-Induced Hypoglycemia in Mouse Retina. PLoS ONE, 11(2), e0150266.

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Purification and Immobilization of Factor H

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Frozen recovered human plasma was obtained from volunteer donors from the San Diego Blood Bank. DEAE-Sephadex and Sephacryl S300 were from GE Healthcare (Little Chalfont, UK). 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Hepes), Polyethylene Glycol 4000 (PEG), (N-morpholino)ethane sulfonic acid (Mes), (N-morpholino)propanesulfonic acid (Mops), Horseradish Peroxidase (HRP), 3,3′,5,5′-tetramethylbenzidine (TMB) and 3-amino-9-ethylcarbazole were from Sigma-Aldrich Chemical Corp. (St. Louis, MO). Luminata forte chemoluminescent HRP detection reagent for ELISA was from EMD/Millipore (Billerica, MA). A murine mAb (clone B-5) directed against the amino terminal modules of human Fibulin-1 was purchased from Santa Cruz Biotechnology (Dallas, TX). Goat anti-rabbit IgG coupled to HRP was purchased from Life Technologies (Carlsbad, CA). Factor H was purified from human plasma similarly to a published method using the following steps: 4–12% PEG precipitation of barium citrate treated plasma, DEAE-Sephadex column chromatography, gel filtration through Sephacryl S300, and column chromatography on dextran sulfate Sepharose [29 (link)]. Factor H-Sepharose was prepared by conjugating Factor H (10 mg/ml) in 0.15 M NaHCO₃ pH 8.2 to Sepharose 6B using the cyanogen bromide reaction [30 ].

DiScipio R.G., Liddington R.C, & Schraufstatter I.U. (2016). Fibulin-1 purification from human plasma using affinity chromatography on Factor H-Sepharose. Protein expression and purification, 121, 118-124.

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