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Dq collagen

Manufactured by Thermo Fisher Scientific
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DQ collagen is a type of collagen-based fluorescent substrate used for the detection and quantification of collagenolytic enzyme activity. It provides a simple and sensitive method for measuring collagenase activity in various biological samples.

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Lab products found in correlation

Enzchek gelatinase collagenase assay kit, by Thermo Fisher Scientific (2 mentions) Dq gelatin, by Thermo Fisher Scientific (2 mentions) Axio observer z1, by Zeiss (1 mentions) Human collagen type 1, by Advanced BioMatrix (1 mentions) Macrophage serum free medium, by Thermo Fisher Scientific (1 mentions) Glomax discover, by Promega (1 mentions) Batimastat, by Merck Group (1 mentions) Su6656, by Santa Cruz Biotechnology (1 mentions) Draq5, by Cell Signaling Technology (1 mentions) Alexa fluor conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Oregon green 488 conjugated gelatin, by Thermo Fisher Scientific (1 mentions) Dasatinib, by Santa Cruz Biotechnology (1 mentions) Timp 1, by Merck Group (1 mentions) Human phospho kinase antibody array, by R&D Systems (1 mentions) Timp2, by Merck Group (1 mentions) Eclipse ti e inverted microscope, by Nikon (1 mentions) Slowfade gold, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 protein labeling kit, by Thermo Fisher Scientific (1 mentions) Gfr matrigel, by BD (1 mentions) Latrunculin b, by Merck Group (1 mentions)

Spelling variants of this product

DQ™ type I collagen (16 citations) DQ-collagen IV (12 citations) DQ-FITC collagen I or IV (3 citations) DQ®-FITC-conjugated type I collagen (2 citations)

11 protocols using dq collagen

1

Quantifying Pericellular Protease Activity

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Analyzing protease-mediated matrix degradation was performed as previously published(16 ) by incorporating 10 μg/mL of DQ collagen (Invitrogen) into polymer solution prior to cell encapsulation. On day 3 DQ collagen was analyzed by fluoresce microscopy (Axio Observer Z1 Zeiss) and measured using fluorescence spectrophotometry at a wavelength of 495 nm excitation and 515 nm emissions (Molecular Devices). High resolution fluorescence and light microscopy images were taken and overlaid to determine the localized protease activity in the cells’ surrounding.
Lewis D.M., Pruitt H., Jain N., Ciccaglione M., McCaffery J.M., Xia Z., Weber K., Eisinger-Mathason T.S, & Gerecht S. (2019). A feedback loop between hypoxia and matrix stress relaxation increases oxygen-axis migration and metastasis in sarcoma. Cancer research, 79(8), 1981-1995.
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2

In Vitro Collagen Cleavage and Remodeling

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To assess collagen cleavage in vitro, cells were incubated for 72 hours in media or media containing 20 μg/mL dye-quenched (DQ) collagen (DQ Collagen, type I from Bovine Skin, Fluorescein Conjugate, D12060) (Invitrogen, Waltham, MA) and fluorescence was measured on a plate reader and normalized to media-only containing wells with and without DQ Collagen. To assess DQ Collagen remodeling during VM formation in vitro, DQ Collagen was added to Matrigel at a concentration of 100 μg/mL, and a VM assay was performed as described above. Brightfield and fluorescence images were acquired, processed (background removal standardized to all images within the same experiment and green false color on fluorescence images), and overlayed in ImageJ.
Pearsall S.M., Williamson S.C., Humphrey S., Hughes E., Morgan D., Marqués F.J., Awanis G., Carroll R., Burks L., Shue Y.T., Bermudez A., Frese K.K., Galvin M., Carter M., Priest L., Kerr A., Zhou C., Oliver T.G., Humphries J.D., Humphries M.J., Blackhall F., Cannell I.G., Pitteri S.J., Hannon G.J., Sage J., Dive C, & Simpson K.L. (2023). Lineage Plasticity in SCLC Generates Non-Neuroendocrine Cells Primed for Vasculogenic Mimicry. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 18(10), 1362-1385.
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3

Collagen and Gelatin Degradation Assay

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Type I collagen and gelatin degradation was assessed using the EnzChek Gelatinase/Collagenase Assay kits (Molecular Probes). Samples were activated with 2 mM of 4-amino-phenyl mercuric acetate (APMA) for 1 hour at 37°C. 80μL of reaction buffer or inhibitor (doxycyline hyclate, Goat anti-human MMP-8 or Mouse anti-human MMP-9) were added with 20μL of either DQ collagen or gelatin (Invitrogen) at a final concentration of 25μg/ml. Activated samples were subsequently added, and activity detected at specified times using a fluorometer (FLUOstar Galaxy).
Ong C.W., Elkington P.T., Brilha S., Ugarte-Gil C., Tome-Esteban M.T., Tezera L.B., Pabisiak P.J., Moores R.C., Sathyamoorthy T., Patel V., Gilman R.H., Porter J.C, & Friedland J.S. (2015). Neutrophil-Derived MMP-8 Drives AMPK-Dependent Matrix Destruction in Human Pulmonary Tuberculosis. PLoS Pathogens, 11(5), e1004917.
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4

Quantifying Type IV Collagen Degradation

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Type IV collagen degradation was assessed using the EnzChek ® Gelatinase/Collagenase Assay kit (Molecular Probes). Samples were activated with 2 mM of 4-amino-phenyl mercuric acetate (APMA) for 1 h at 37 °C. Eighty microliters of reaction buffer or inhibitor (mouse anti-human MMP-9) were added with 20 μL of DQ collagen (Invitrogen) at a final concentration of 25 μg/ml. The activated samples were subsequently added, and activity was detected at specified times using a fluorometer (FLUOstar Galaxy).
Ong C.W., Pabisiak P.J., Brilha S., Singh P., Roncaroli F., Elkington P.T, & Friedland J.S. (2017). Complex regulation of neutrophil-derived MMP-9 secretion in central nervous system tuberculosis. Journal of Neuroinflammation, 14, 31.
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5

In Vitro Collagen Cleavage and Remodeling

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To assess collagen cleavage in vitro, cells were incubated for 72 hours in media or media containing 20 μg/mL dye-quenched (DQ) collagen (DQ Collagen, type I from Bovine Skin, Fluorescein Conjugate, D12060) (Invitrogen, Waltham, MA) and fluorescence was measured on a plate reader and normalized to media-only containing wells with and without DQ Collagen. To assess DQ Collagen remodeling during VM formation in vitro, DQ Collagen was added to Matrigel at a concentration of 100 μg/mL, and a VM assay was performed as described above. Brightfield and fluorescence images were acquired, processed (background removal standardized to all images within the same experiment and green false color on fluorescence images), and overlayed in ImageJ.
Cannon D.S, & Allen S.N. (2000). A Comparison of the Effects of Computer and Manual Reminders on Compliance with a Mental Health Clinical Practice Guidelines. Journal of the American Medical Informatics Association : JAMIA, 7(2), 196-203.
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6

Extracellular Matrix Degradation Assay

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For analysis of extracellular matrix degradation, microspheres were generated from a solution of 3% Alginate (Pronova UP MVG alginate, Nova Matrix, Norway), 1 mg/ml of human collagen type I (Advanced BioMatrix, San Diego, California) and 100 µg/ml of DQ collagen (Invitrogen, Paisley, UK). Microspheres were then placed in macrophage serum free medium (Life Technologies) and incubated at 37°C. Fluorescence was read on a GloMax Discover (Promega) at an absorption maxima of 495 nm and fluorescence emission maxima of 515 nm.
Tezera L.B., Bielecka M.K., Chancellor A., Reichmann M.T., Shammari B.A., Brace P., Batty A., Tocheva A., Jogai S., Marshall B.G., Tebruegge M., Jayasinghe S.N., Mansour S, & Elkington P.T. (2017). Dissection of the host-pathogen interaction in human tuberculosis using a bioengineered 3-dimensional model. eLife, 6, e21283.
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7

Trophoblast cell colony analysis

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Culture medium of trophoblast cell colony on the fibroblast feeder layer or in the absence of feeder cells was changed to DMEM (+FBS) containing dye-quenched (DQ) collagen (50-fold dilution) (Invitrogen, USA) [14 (link),15 (link)]. After incubation for 2 h, the cell colonies were once washed with DMEM (−FBS). And the medium was changed to DMEM (−FBS) containing 4 to 40 μM of 12-KETE. Chromatic images were observed using a fluorescence microscope (IX70; OLYMPUS, Japan).
, & Kamada H. (2022). 12-Oxoeicosatetraenoic acid, a candidate signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells. Animal Bioscience, 36(3), 429-440.
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8

Integrin-mediated Cell Adhesion Assays

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TIMP1, TIMP2, and batimastat were from Sigma-Aldrich. Rabbit anti-MMP14 (hinge region) and mouse anti–integrin β1 antibodies were from Abcam (Cambridge, MA). Mouse anti-cortactin antibody was from Millipore (Billerica, MA). Rabbit anti–p-c-Src (Tyr-419) antibody and Src family kinase inhibitors Dasatinib and SU6656 were from Santa Cruz Biotechnology (Santa Cruz, CA). DQ collagen, Oregon Green 488–conjugated gelatin, mouse anti–cadherin-11, mouse anti–α-catenin, and mouse anti–β-catenin antibodies, Alexa Fluor–conjugated anti-mouse, anti-rat, and anti-rabbit secondary antibodies, and Alexa Fluor–conjugated phalloidin were from Invitrogen. Mouse anti–p120-catenin antibody was from BD Biosciences (San Jose, CA). Mouse anti–cadherin-11 (3H10) was previously raised in this laboratory (Valencia et al., 2004 (link)). DRAQ5 was from Cell Signaling (Danvers, MA). Human phosphokinase antibody array was from R&D Systems (Minneapolis, MN).
Gu Z., Liu F., Tonkova E.A., Lee S.Y., Tschumperlin D.J, & Brenner M.B. (2014). Soft matrix is a natural stimulator for cellular invasiveness. Molecular Biology of the Cell, 25(4), 457-469.
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9

In-situ Zymography for MMP Activity

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MMP activity was measured using in-situ zymography as previously described [83 (link)]. 5-μm paraffin embedded sections were wax-cleared and rehydrated. As per manufacturers instructions, skin sections were incubated with DQ Gelatin or DQ Collagen 1:200 (Thermofisher) in reaction buffer (50 mM Tris, 150 mM NaCI, 5 mM CaCl2, 0.2 mM NaN3) for 2 hours. Control sections were treated with reaction buffer alone (no DQ Gelatin or DQ Collagen). Sections were then treated with Sudan black, counterstained with DAPI and mounted with slowfade gold (Thermofisher). Using a motorized stage, 20x fluorescence images of the entire skin section were acquired using a Nikon Eclipse Ti-E inverted microscope.
Govindaraju P., Todd L., Shetye S., Monslow J, & Puré E. (2018). CD44-dependent inflammation, fibrogenesis, and collagenolysis regulates extracellular matrix remodeling and tensile strength during cutaneous wound healing. Matrix biology : journal of the International Society for Matrix Biology, 75-76, 314-330.
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10

Immunofluorescence Microscopy with Cytoskeletal Markers

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The anti-MAP2 antibody was acquired from Antibodies Online (ABIN372661; used at 1:1,000 for immunofluorescence). The Alexa Fluor 488 Phalloidin (A12379), used to stain filamentous actin, and DAPI (D1306) were purchased from Life Technologies. Bovine plasma FN was purchased from Merck (341631). DMSO and latrunculin B (L5288-1MG) were obtained from Sigma-Aldrich. GFR Matrigel was bought from BD Biosciences (354230). PureCol EZ Gel (fibrillar collagen I, concentration 5 mg/ml) was provided by Advanced BioMatrix. DQ collagen (type I collagen from bovine skin, fluorescein conjugate; D12060) was provided by Thermo Fisher Scientific. mEmerald-Lifeact-7 was a gift from M. Davidson (plasmid 54148; Addgene). psPAX2 and pMD2.G were gifts from D. Trono (École polytechnique fédérale de Lausanne, Lausanne, Switzerland; plasmids 12260 and 12259; Addgene). pCDH-LifeAct-mRFP was a gift from P. Caswell. Full-length bovine FN was labeled with Alexa Fluor 568 using an Alexa Fluor 568 Protein Labeling kit (A10238; Thermo Fisher Scientific) according to the manufacturer’s instructions.
Jacquemet G., Paatero I., Carisey A.F., Padzik A., Orange J.S., Hamidi H, & Ivaska J. (2017). FiloQuant reveals increased filopodia density during breast cancer progression. The Journal of Cell Biology, 216(10), 3387-3403.
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