Collagen 1 rat protein

Manufactured by Thermo Fisher Scientific

Collagen I Rat Protein is a purified protein derived from rat collagen type I. It is a structural protein that provides strength and support to various tissues in the body. This product is suitable for use in cell culture, tissue engineering, and biochemical applications.

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Lab products found in correlation

N acetyl l cysteine nac, by Merck Group (1 mentions) Recombinant human tnf, by R&D Systems (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Penicilin streptomycin, by Thermo Fisher Scientific (1 mentions) Deferoxamine dfo, by Merck Group (1 mentions) Axiovert 200m, by Zeiss (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Filipin 3, by Merck Group (1 mentions) C4742 80 12ag camera, by Hamamatsu Photonics (1 mentions) In cell analyzer 6000, by GE Healthcare (1 mentions) Viewplate 96 black, by PerkinElmer (1 mentions)

Close spelling variants of this product

Collagen I (143 citations) Rat collagen I (5 citations) Collagen I from rat protein solution (2 citations)

4 protocols using collagen 1 rat protein

Generation of Genetically Engineered Mouse Organoids

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Three-dimensional (3D) mouse organoids were created from PbCre negative mice with floxed alleles (Pten^f/f, Pten^f/f; LSL-MYCN+, Pten^f/f; Rb1^f/f, Pten^f/f; LSL-MYCN+; Rb1^f/f) as described above. Newly created 3D organoids were passaged three times before being plated in 2D cell culture. Organoids were dissociated as described above and dissociated cells were plated on 1% Collagen I Rat Protein (Gibco, A1048301) coated cell culture flasks. After an additional two passages, cells were seeded at 250,000 cells/well in a 6-well plate. The following day, cells were infected at an MOI of 500. We used recombinant empty vector (Vector Biolabs, 1660) as a control and Cre-expressing (Vector Biolabs, 1774) human adenoviruses type 5 expressing Red Fluorescent Protein (RFP) and a Cre recombinase under the control of the CMV promoter. The following day, cells were washed once in PBS and new media was added. Cells were propagated for an additional few passages before growing cells as 3D organoids. The resulting P, PN, and PRN organoids were named according to the adenovirus used (e.g., the empty vector: P^Con, PN^Con, PRN^Con; or Cre-expressing adenovirus: P^Cre, PN^Cre, PRN^Cre).

Brady N.J., Bagadion A.M., Singh R., Conteduca V., Van Emmenis L., Arceci E., Pakula H., Carelli R., Khani F., Bakht M., Sigouros M., Bareja R., Sboner A., Elemento O., Tagawa S., Nanus D.M., Loda M., Beltran H., Robinson B, & Rickman D.S. (2021). Temporal evolution of cellular heterogeneity during the progression to advanced AR-negative prostate cancer. Nature Communications, 12, 3372.

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Evaluating G6Pase in HepG2 cells

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HepG2 cells were grown in low glucose (1g/L) DMEM (GIBCO® Ref: 21885-108), 10% heat inactivated FBS, 1% Penicilin/Streptomycin (GIBCO® Ref: 15140-122). Before seeding, plates were coated with collagen I rat protein (GIBCO, ThermoFisher Scientific; 5μg/ml in 20mM acetic) acid. At 100% confluence cells were treated with heme (Frontier Scientifc; 40 μM, 1 hr) in HBSS, human recombinant TNF (R&D Systems Europe Ref: 210-TA-20) was added in media (50 ng/mL; 4 hr) and cells were collected. When indicated cells were pre-treated with N-acetyl-L-cysteine (NAC; Sigma, Ref: A9165; 7.5mM in media; 3 hr) before and during heme treatment in HBSS and subsequently throughout the experiment. When indicated cells were pre-treated Deferoxamine (DFO, Sigma Ref: D9533; 60 μM; 16 hr) before and during heme treatment in HBSS and subsequently throughout the experiment. At 80%–90% of confluence, cells were transduced at 50 or 100 pfu with LacZ, G6pc1 (Vector Biolabs; Ref: SKU#: ADV-259766), FTH and FTH^m Rec.Ad. (Berberat et al., 2003 (link), Gozzelino et al., 2012 (link)). Medium was replaced 24 hr after Rec.Ad. transduction. G6Pase mRNA, protein and activity were analyzed 48 or 72 hr after Rec.Ad. transduction, as described below.

Weis S., Carlos A.R., Moita M.R., Singh S., Blankenhaus B., Cardoso S., Larsen R., Rebelo S., Schäuble S., Del Barrio L., Mithieux G., Rajas F., Lindig S., Bauer M, & Soares M.P. (2017). Metabolic Adaptation Establishes Disease Tolerance to Sepsis. Cell, 169(7), 1263-1275.e14.

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Visualizing Cholesterol Accumulation in HAP1 Cells

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Coverslips were incubated for 1 h at 37°C in 1 mL of 1:30 solution of collagen I rat protein (Thermo Fisher Scientific) to 1× PBS. The collagen solution was then aspirated, and 250,000 HAP1 cells were seeded onto the coverslips in 12-well plate 24 h before staining. Cells were washed three times with 1× PBS and then fixed in 4% paraformaldehyde in 1× PBS for 30 min at room temperature. After fixation, cells were washed three times in 1× PBS and then stained with 50 μg/mL of filipin III (Sigma-Aldrich) for 1 h at room temperature. Coverslips were then washed three times in 1× PBS and mounted onto a slide using ProLong Gold Antifade Mountant (Thermo Fisher Scientific). Cells were visualized using the Zeiss Axiovert 200M epifluorescent microscope, and images were captured with a Hamamatsu C4742-80-12AG camera.

Erwood S., Brewer R.A., Bily T.M., Maino E., Zhou L., Cohn R.D, & Ivakine E.A. (2019). Modeling Niemann–Pick disease type C in a human haploid cell line allows for patient variant characterization and clinical interpretation. Genome Research, 29(12), 2010-2019.

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Quantifying Fluorescent Protein Expression in HepG2 Cells

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PerkinElmer ViewPlate-96 Black plates were coated with 2% collagen for 10 min at RT (100% collagen I rat protein was obtained from Thermo Fisher Scientific and diluted to 2% in UltraPure water). Wells were washed with PBS, and 10,000 HepG2 cells were plated per well. The following day, cells were transfected with indicated plasmids (100 ng/well) using Lipofectamine 3000. Twenty-four hours after transfection, compounds were added at the indicated concentrations in phenol red–free DMEM supplemented with 5% charcoal/dextran-treated FBS. The final DMSO concentration was 0.25%. After an additional 24 h, cells were washed with PBS, crosslinked with 4% formaldehyde in PBS at RT for 20 min, washed 3x for 5 min with PBS, permeabilized with 0.1% Triton X-100 in PBS for 10 min, washed 3x for 5 min with PBS, blocked with 5% BSA in PBST (PBS, 0.1% Tween 20) for 1 h, rinsed 1x with PBS, stained with mouse anti-FLAG M2 (1:500 dilution) in PBST with 1% BSA at RT for 1 h, washed 3x for 5 min with PBST, stained with goat anti-mouse Alexa Fluor 488 (1:500 dilution) in PBST with 1% BSA at RT for 1 h, stained with 1 μg/mL DAPI in PBS for 15 min, washed 3x for 5 min with PBST, and stored in PBS. Images were taken with an IN CELL Analyzer 6000 (GE Healthcare) using a 20× objective. CellProfiler was used for quantification [46 (link)–48 (link)].

Huber A.D., Wright W.C., Lin W., Majumder K., Low J.A., Wu J., Buchman C.D., Pintel D.J, & Chen T. (2020). Mutation of a single amino acid of pregnane X receptor switches an antagonist to agonist by altering AF-2 helix positioning. Cellular and molecular life sciences : CMLS, 78(1), 317-335.

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