collagenase i  (Worthington Biochemical)


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    Structured Review

    Worthington Biochemical collagenase i
    Collagenase I, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase i/product/Worthington Biochemical
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase i - by Bioz Stars, 2022-09
    99/100 stars

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    Worthington Biochemical collagenase type 1
    AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar <t>type</t> 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.
    Collagenase Type 1, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase type 1/product/Worthington Biochemical
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase type 1 - by Bioz Stars, 2022-09
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    95
    Worthington Biochemical collagenase type i
    Differences in the capacity of myogenic cells from slow- and fast-type muscles. a Freshly isolated intact myofibers from EDL or <t>SOL</t> muscle of wild-type mice were stained against Pax7 (red) and with DAPI (blue). Typical Pax7(+) and DAPI(+) cells are shown (arrowheads). The number of Pax7(+) satellite cells per fiber was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. b Myofibers isolated from SOL or EDL muscle were cultured under growth conditions for 48 h. The activated satellite cells that migrated from each muscle fiber (arrowheads) were counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. c Myoblasts from SOL or TA muscle were cultured in growth medium with EdU. After staining with EdU (green) and DAPI (blue), the proportion of EdU(+) cells among the total nuclei was counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. d Myoblasts from SOL or TA muscle were cultured in differentiation medium for 4 days, and then myotubes and nuclei were stained against MyHC (green) and with DAPI (blue). The proportion of MyHC(+) cells among the total nuclei was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. e Myotubes of TA- and SOL-derived myoblasts were stained against Myh7 (MyHC <t>type</t> I slow; green) and with DAPI (blue). Scale bar = 200 µm. f The expression levels of Myh7 , Myh2 , Myh1 , and Myh4 in myotubes were quantified by qPCR. The expression values were normalized to GAPDH expression and are presented as mean ± SE ( n = 4). g TA- or SOL-MBs derived from GFP mouse muscles were injected into TA muscles of NOD/ scid mice. Typical phase image (left) and GFP expression (right) are presented. Scale bar = 2 mm. h Longitudinal sections of engrafted TA muscles stained against slow-type myosin heavy chain (red) and laminin-α2 (gray), and with DAPI (blue). Scale bar = 500 µm. i OCR in SOL- or TA-derived myotubes was measured after treatment with oligomycin and FCCP. Values are presented as mean ± SE ( n = 3). j Basal OCR and SRC of myotubes were quantified. Values are presented as mean ± SE ( n = 3). k The expression levels of PGC-1 α, Nrf1 , and Tfam in TA- and SOL-MTs were quantified by qPCR. Values are presented as mean ± SE ( n = 3)
    Collagenase Type I, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
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    AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar type 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A lung tropic AAV vector improves survival in a mouse model of surfactant B deficiency

    doi: 10.1038/s41467-020-17577-8

    Figure Lengend Snippet: AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar type 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.

    Article Snippet: Following perfusion, an enzyme mix containing 30 U neutral protease (Worthington; LS02104), 2500 U Collagenase I (Worthington; LS004196), 10 μg DNase I (Sigma; D5025-150KU) in DPBS with Mg2+ and Ca2+ was instilled into the lungs.

    Techniques: Staining, Plasmid Preparation, Mouse Assay, Flow Cytometry, Transgenic Assay, Expressing, Two Tailed Test

    Tm4sf1 -marked subpopulation of endothelial cells expresses a distinct transcriptomic signature relevant to PAH. (A-F) Uniform manifold approximation and projection (UMAP) plots showing clustering of 758 endothelial cell transcriptomes from 18 rats lungs with individual cells colored by (A) subpopulation, (B) disease condition ( n = 6/group), and expression levels of the following marker genes: (C) Car4 for endothelial capillary (EC), (D) Nostrin for endothelial arterial type 1 (EA), (E) Tm4sf1 for endothelial arterial type 2 (EA2), and (F) Sox17 as a known arterial marker expressed in both EA1 and EA2. Expression levels are represented as log normalized counts. (G) Volcano plot showing DEGs in 255 EA2 cells compared to 454 EA1 cells where x-axis represents MAST z-scores and y-axis indicates - log 10 ( P ). Significant upregulated (z > 0) or downregulated (z

    Journal: bioRxiv

    Article Title: A Tm4sf1-Marked Subpopulation of Endothelial Stem/Progenitor Cells Identified by Lung Single-Cell Omics of Pulmonary Arterial Hypertension

    doi: 10.1101/2022.01.09.475566

    Figure Lengend Snippet: Tm4sf1 -marked subpopulation of endothelial cells expresses a distinct transcriptomic signature relevant to PAH. (A-F) Uniform manifold approximation and projection (UMAP) plots showing clustering of 758 endothelial cell transcriptomes from 18 rats lungs with individual cells colored by (A) subpopulation, (B) disease condition ( n = 6/group), and expression levels of the following marker genes: (C) Car4 for endothelial capillary (EC), (D) Nostrin for endothelial arterial type 1 (EA), (E) Tm4sf1 for endothelial arterial type 2 (EA2), and (F) Sox17 as a known arterial marker expressed in both EA1 and EA2. Expression levels are represented as log normalized counts. (G) Volcano plot showing DEGs in 255 EA2 cells compared to 454 EA1 cells where x-axis represents MAST z-scores and y-axis indicates - log 10 ( P ). Significant upregulated (z > 0) or downregulated (z

    Article Snippet: For the second (n = 3/group for scRNA-seq) and third (n = 4/group for FACS) sets of rats, minced lungs were incubated with collagenase I (Worthington, cat. no. LS004194) at 1 mg/ml in DMEM + 10% FBS in a 37 °C water bath for 60 min, with trituration every 5 minutes ( – )(3).

    Techniques: Expressing, Marker

    Differences in the capacity of myogenic cells from slow- and fast-type muscles. a Freshly isolated intact myofibers from EDL or SOL muscle of wild-type mice were stained against Pax7 (red) and with DAPI (blue). Typical Pax7(+) and DAPI(+) cells are shown (arrowheads). The number of Pax7(+) satellite cells per fiber was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. b Myofibers isolated from SOL or EDL muscle were cultured under growth conditions for 48 h. The activated satellite cells that migrated from each muscle fiber (arrowheads) were counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. c Myoblasts from SOL or TA muscle were cultured in growth medium with EdU. After staining with EdU (green) and DAPI (blue), the proportion of EdU(+) cells among the total nuclei was counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. d Myoblasts from SOL or TA muscle were cultured in differentiation medium for 4 days, and then myotubes and nuclei were stained against MyHC (green) and with DAPI (blue). The proportion of MyHC(+) cells among the total nuclei was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. e Myotubes of TA- and SOL-derived myoblasts were stained against Myh7 (MyHC type I slow; green) and with DAPI (blue). Scale bar = 200 µm. f The expression levels of Myh7 , Myh2 , Myh1 , and Myh4 in myotubes were quantified by qPCR. The expression values were normalized to GAPDH expression and are presented as mean ± SE ( n = 4). g TA- or SOL-MBs derived from GFP mouse muscles were injected into TA muscles of NOD/ scid mice. Typical phase image (left) and GFP expression (right) are presented. Scale bar = 2 mm. h Longitudinal sections of engrafted TA muscles stained against slow-type myosin heavy chain (red) and laminin-α2 (gray), and with DAPI (blue). Scale bar = 500 µm. i OCR in SOL- or TA-derived myotubes was measured after treatment with oligomycin and FCCP. Values are presented as mean ± SE ( n = 3). j Basal OCR and SRC of myotubes were quantified. Values are presented as mean ± SE ( n = 3). k The expression levels of PGC-1 α, Nrf1 , and Tfam in TA- and SOL-MTs were quantified by qPCR. Values are presented as mean ± SE ( n = 3)

    Journal: Cell Death and Differentiation

    Article Title: Tbx1 regulates inherited metabolic and myogenic abilities of progenitor cells derived from slow- and fast-type muscle

    doi: 10.1038/s41418-018-0186-4

    Figure Lengend Snippet: Differences in the capacity of myogenic cells from slow- and fast-type muscles. a Freshly isolated intact myofibers from EDL or SOL muscle of wild-type mice were stained against Pax7 (red) and with DAPI (blue). Typical Pax7(+) and DAPI(+) cells are shown (arrowheads). The number of Pax7(+) satellite cells per fiber was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. b Myofibers isolated from SOL or EDL muscle were cultured under growth conditions for 48 h. The activated satellite cells that migrated from each muscle fiber (arrowheads) were counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. c Myoblasts from SOL or TA muscle were cultured in growth medium with EdU. After staining with EdU (green) and DAPI (blue), the proportion of EdU(+) cells among the total nuclei was counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. d Myoblasts from SOL or TA muscle were cultured in differentiation medium for 4 days, and then myotubes and nuclei were stained against MyHC (green) and with DAPI (blue). The proportion of MyHC(+) cells among the total nuclei was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. e Myotubes of TA- and SOL-derived myoblasts were stained against Myh7 (MyHC type I slow; green) and with DAPI (blue). Scale bar = 200 µm. f The expression levels of Myh7 , Myh2 , Myh1 , and Myh4 in myotubes were quantified by qPCR. The expression values were normalized to GAPDH expression and are presented as mean ± SE ( n = 4). g TA- or SOL-MBs derived from GFP mouse muscles were injected into TA muscles of NOD/ scid mice. Typical phase image (left) and GFP expression (right) are presented. Scale bar = 2 mm. h Longitudinal sections of engrafted TA muscles stained against slow-type myosin heavy chain (red) and laminin-α2 (gray), and with DAPI (blue). Scale bar = 500 µm. i OCR in SOL- or TA-derived myotubes was measured after treatment with oligomycin and FCCP. Values are presented as mean ± SE ( n = 3). j Basal OCR and SRC of myotubes were quantified. Values are presented as mean ± SE ( n = 3). k The expression levels of PGC-1 α, Nrf1 , and Tfam in TA- and SOL-MTs were quantified by qPCR. Values are presented as mean ± SE ( n = 3)

    Article Snippet: Isolation and culture of single muscle fibersThe EDL, TA, and SOL muscles isolated from mice were digested with 2 mg/ml collagenase type I (Worthington Biochemical), as previously described [ ], and the isolated muscle fibers were suspended and cultured in growth medium (DMEM containing 15% horse serum [Invitrogen] with 0.5% chick embryo extract [Gemini Bio-Products, West Sacramento, CA] and penicillin–streptomycin) at 37 °C [ ].

    Techniques: Isolation, Mouse Assay, Staining, Cell Culture, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Pyrolysis Gas Chromatography