non parenchymal cells npcs (Worthington Biochemical)

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Collagenase Type 2

Description:

Prepared to contain higher clostripain activity Suggested for bone heart liver thyroid and salivary primary cell isolation Supplied as a dialyzed lyophilized powder

Catalog Number:

ls004174

Price:

Size:

100 mg

Source:

Clostridium histolyticum

Cas Number:

9001.12.1

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Worthington Biochemical non parenchymal cells npcs
Microscope images of <t>HCs</t> and <t>NPCs</t> in culture. These pictures depict representative areas of untreated, PB in vitro as well as in vivo treated HCs and NPCs in culture extracted from microscopic images of equal magnification.

Microscope images of <t>HCs</t> and <t>NPCs</t> in culture. These pictures depict representative areas of untreated, PB in vitro as well as in vivo treated HCs and NPCs in culture extracted from microscopic images of equal magnification.

Prepared to contain higher clostripain activity Suggested for bone heart liver thyroid and salivary primary cell isolation Supplied as a dialyzed lyophilized powder
https://www.bioz.com/result/non parenchymal cells npcs/product/Worthington Biochemical
Average 85 stars, based on 2919 article reviews
Price from $9.99 to $1999.99

non parenchymal cells npcs - by Bioz Stars, 2020-05

85/100 stars

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Images

1) Product Images from "Phenobarbital Induces Alterations in the Proteome of Hepatocytes and Mesenchymal Cells of Rat Livers"

Article Title: Phenobarbital Induces Alterations in the Proteome of Hepatocytes and Mesenchymal Cells of Rat Livers

Journal: PLoS ONE

doi: 10.1371/journal.pone.0076137

Microscope images of HCs and NPCs in culture. These pictures depict representative areas of untreated, PB in vitro as well as in vivo treated HCs and NPCs in culture extracted from microscopic images of equal magnification.

Figure Legend Snippet: Microscope images of HCs and NPCs in culture. These pictures depict representative areas of untreated, PB in vitro as well as in vivo treated HCs and NPCs in culture extracted from microscopic images of equal magnification.

Techniques Used: Microscopy, In Vitro, In Vivo

$Proteome alterations induced by in vitro treatment of primary cells. Part A) shows schematic representations of a cell and her three sub-compartments, namely the supernatant, the cytoplasm and the nucleus. The intensity of red represents the degree of amount of the selected protein found in the respective compartment in contrast to the other experiments. The higher intensity of red corresponds to a higher occurrence. This allows an easy comparison of the expression levels of a protein in different experimental setups. NPCs induce the secretion of IL-1beta and TNF-alpha upon inflammatory stimulation with LPS. In vitro treatment with PB induced coronin-7 and ADP-ribosyl cyclase 1, which both are also induced by in vivo treatment. The expression of Hsp90, a stress response related protein, was increased upon LPS and PB treatment. Prostaglandin, a protein involved in promotion of proliferation in normal and preneoplastic cells, was induced upon LPS and in vivo PB treatment. HCs respond hardly to the in vitro treatment with PB. Treatment with IL-6 specifically induced the acute phase protein T-kininogen-2. UDP-glucuronosyltransferase 2B37 and the chaperone peptidyl-prolyl cis-trans isomerase D were induced by both in vitro stimulation experiments as well as by the in vivo treatment with PB. Carbamoyl-phosphate synthase is part of the urea cycle and has to be found in all four categories. Proteins in NPC: (1) O35828 Coronin-7, (2) P16599 Tumor necrosis factor, (3) P34058 Heat shock protein HSP 90-beta, (4) Q63264 Interleukin-1 beta, (5) Q63921 Prostaglandin G/H synthase 1, (6) Q64244 ADP-ribosyl cyclase 1. Proteins in HC: (1) P07756 Carbamoyl-phosphate synthase [ammonia], (2) P08932 T-kininogen 2, (3) P19488 UDP-glucuronosyltransferase 2B37, (4) Q6DGG0 Peptidyl-prolyl cis-trans isomerase D. Part B) demonstrates the distribution of distinct proteins within the three fractions, supernatant, cytoplasm and nuclear protein fractions, underneath the respective treatment of the cells, which gives an overview of the responsiveness of the cells. Abbr.: SN –proteome of the supernatant, Cyt – proteome of the cytoplasm, NE – proteome of the nuclear extract.$
Figure Legend Snippet: Proteome alterations induced by in vitro treatment of primary cells. Part A) shows schematic representations of a cell and her three sub-compartments, namely the supernatant, the cytoplasm and the nucleus. The intensity of red represents the degree of amount of the selected protein found in the respective compartment in contrast to the other experiments. The higher intensity of red corresponds to a higher occurrence. This allows an easy comparison of the expression levels of a protein in different experimental setups. NPCs induce the secretion of IL-1beta and TNF-alpha upon inflammatory stimulation with LPS. In vitro treatment with PB induced coronin-7 and ADP-ribosyl cyclase 1, which both are also induced by in vivo treatment. The expression of Hsp90, a stress response related protein, was increased upon LPS and PB treatment. Prostaglandin, a protein involved in promotion of proliferation in normal and preneoplastic cells, was induced upon LPS and in vivo PB treatment. HCs respond hardly to the in vitro treatment with PB. Treatment with IL-6 specifically induced the acute phase protein T-kininogen-2. UDP-glucuronosyltransferase 2B37 and the chaperone peptidyl-prolyl cis-trans isomerase D were induced by both in vitro stimulation experiments as well as by the in vivo treatment with PB. Carbamoyl-phosphate synthase is part of the urea cycle and has to be found in all four categories. Proteins in NPC: (1) O35828 Coronin-7, (2) P16599 Tumor necrosis factor, (3) P34058 Heat shock protein HSP 90-beta, (4) Q63264 Interleukin-1 beta, (5) Q63921 Prostaglandin G/H synthase 1, (6) Q64244 ADP-ribosyl cyclase 1. Proteins in HC: (1) P07756 Carbamoyl-phosphate synthase [ammonia], (2) P08932 T-kininogen 2, (3) P19488 UDP-glucuronosyltransferase 2B37, (4) Q6DGG0 Peptidyl-prolyl cis-trans isomerase D. Part B) demonstrates the distribution of distinct proteins within the three fractions, supernatant, cytoplasm and nuclear protein fractions, underneath the respective treatment of the cells, which gives an overview of the responsiveness of the cells. Abbr.: SN –proteome of the supernatant, Cyt – proteome of the cytoplasm, NE – proteome of the nuclear extract.

Techniques Used: In Vitro, Expressing, In Vivo

Distribution of distinct proteins, when comparing controls with PB-treatment from the in vitro and in vivo sample pools, respectively. This figure demonstrates the distribution of distinct proteins found in HCs and NPCs during the pooled A) in vitro and B) in vivo experiments, while including only proteins found with at least 2 peptides. The up- and down-regulation of proteins were neglected in this qualitative comparison.

Figure Legend Snippet: Distribution of distinct proteins, when comparing controls with PB-treatment from the in vitro and in vivo sample pools, respectively. This figure demonstrates the distribution of distinct proteins found in HCs and NPCs during the pooled A) in vitro and B) in vivo experiments, while including only proteins found with at least 2 peptides. The up- and down-regulation of proteins were neglected in this qualitative comparison.

Techniques Used: In Vitro, In Vivo

2) Product Images from "Fli1-haploinsufficient dermal fibroblasts promote skin-localized transdifferentiation of Th2-like regulatory T cells"

Article Title: Fli1-haploinsufficient dermal fibroblasts promote skin-localized transdifferentiation of Th2-like regulatory T cells

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-018-1521-3

T helper type 2 cell (Th2)-like regulatory T cells (Tregs) are induced by coculture with Fli1 +/− dermal fibroblasts through interleukin (IL)-33. a Evaluation by flow cytometry of Th1-, Th2-, and Th17-like Treg induction by coculture with wild-type (WT) and Fli1 +/− dermal fibroblasts ( n = 6). b Evaluation by flow cytometry of the effect of IL-33-neutralizing antibody on Tregs cocultured with Fli1 +/− dermal fibroblasts ( n = 6). Representative plots of interferon (IFN)-γ-, IL-4-, and IL-17A-positive Tregs are shown in right upper panels of ( a ) and ( b ). Gating strategy for identification of CD4 + FoxP3 + Tregs is shown in the leftmost panels of ( a ) and ( b ). In each graph, the relative value compared with the control group is expressed as mean ± SEM. AU Arbitrary units, Fli1 Friend leukemia virus integration 1

Figure Legend Snippet: T helper type 2 cell (Th2)-like regulatory T cells (Tregs) are induced by coculture with Fli1 +/− dermal fibroblasts through interleukin (IL)-33. a Evaluation by flow cytometry of Th1-, Th2-, and Th17-like Treg induction by coculture with wild-type (WT) and Fli1 +/− dermal fibroblasts ( n = 6). b Evaluation by flow cytometry of the effect of IL-33-neutralizing antibody on Tregs cocultured with Fli1 +/− dermal fibroblasts ( n = 6). Representative plots of interferon (IFN)-γ-, IL-4-, and IL-17A-positive Tregs are shown in right upper panels of ( a ) and ( b ). Gating strategy for identification of CD4 + FoxP3 + Tregs is shown in the leftmost panels of ( a ) and ( b ). In each graph, the relative value compared with the control group is expressed as mean ± SEM. AU Arbitrary units, Fli1 Friend leukemia virus integration 1

Techniques Used: Flow Cytometry, Cytometry

3) Product Images from "Fli1-haploinsufficient dermal fibroblasts promote skin-localized transdifferentiation of Th2-like regulatory T cells"

Article Title: Fli1-haploinsufficient dermal fibroblasts promote skin-localized transdifferentiation of Th2-like regulatory T cells

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-018-1521-3

Techniques Used: Flow Cytometry, Cytometry

4) Product Images from "Phenobarbital Induces Alterations in the Proteome of Hepatocytes and Mesenchymal Cells of Rat Livers"

Article Title: Phenobarbital Induces Alterations in the Proteome of Hepatocytes and Mesenchymal Cells of Rat Livers

Journal: PLoS ONE

doi: 10.1371/journal.pone.0076137

Techniques Used: Microscopy, In Vitro, In Vivo

Techniques Used: In Vitro, Expressing, In Vivo

Techniques Used: In Vitro, In Vivo

5) Product Images from "Fluid Shear Stress Alters the Hemostatic Properties of Endothelial Outgrowth Cells"

Article Title: Fluid Shear Stress Alters the Hemostatic Properties of Endothelial Outgrowth Cells

Journal: Tissue Engineering. Part A

doi: 10.1089/ten.tea.2010.0290

Thrombomodulin, nitric oxide synthase 3 (eNOS), tissue factor pathway inhibitor ( TFPI ), tissue factor ( F3 ), and vWF gene expression in ECs and EOCs was determined by quantitative real-time reverse transcriptase–polymerase chain reaction. mRNA

Figure Legend Snippet: Thrombomodulin, nitric oxide synthase 3 (eNOS), tissue factor pathway inhibitor ( TFPI ), tissue factor ( F3 ), and vWF gene expression in ECs and EOCs was determined by quantitative real-time reverse transcriptase–polymerase chain reaction. mRNA

Techniques Used: Expressing, Polymerase Chain Reaction

6) Product Images from "A rigorous method to enrich for exosomes from brain tissue"

Article Title: A rigorous method to enrich for exosomes from brain tissue

Journal: Journal of Extracellular Vesicles

doi: 10.1080/20013078.2017.1348885

$Schematic of the exosome isolation protocol from solid brain tissue. Fresh frozen (−80°C) human frontal cortex was sliced with a razor blade on ice while frozen to generate 1–2 cm long, 2–3 mm wide sections. The cut sections are dissociated while partially frozen in 75 U/ml of collagenase type 3 in Hibernate-E at 37°C for a total of 20 min. The tissue is returned to ice immediately after incubation and protease and phosphatase inhibitors are added. The tissue is spun at 300 × g for 5 min at 4°C (pellet is used as the brain homogenate + collagenase control), the supernatant transferred to a fresh tube, spun at 2000 × g for 10 min at 4°C, then at 10,000 × g for 30 min at 4°C. The EV-containing supernatant is overlaid on a triple sucrose cushion (0.6 M, 1.3 M, 2.5 M) and ultracentrifuged for 3 h at 180,000 × g to separate vesicles based on density. The top of the gradient is discarded and fractions designated 1, 2 and 3 are collected and the refractive index is measured. Each fraction is further ultracentrifuged at 100,000 × g to pellet the vesicles contained in each fraction. Each preparation is validated by a combination of techniques including electron microscopy and RNA and protein analysis. Note – some tissue samples will not be amenable to this method. Post-mortem delay, storage time and the number of freeze-thaw cycles will negatively impact on tissue quality and result in contamination of the fractions with cellular debris and non-exosome vesicles.$
Figure Legend Snippet: Schematic of the exosome isolation protocol from solid brain tissue. Fresh frozen (−80°C) human frontal cortex was sliced with a razor blade on ice while frozen to generate 1–2 cm long, 2–3 mm wide sections. The cut sections are dissociated while partially frozen in 75 U/ml of collagenase type 3 in Hibernate-E at 37°C for a total of 20 min. The tissue is returned to ice immediately after incubation and protease and phosphatase inhibitors are added. The tissue is spun at 300 × g for 5 min at 4°C (pellet is used as the brain homogenate + collagenase control), the supernatant transferred to a fresh tube, spun at 2000 × g for 10 min at 4°C, then at 10,000 × g for 30 min at 4°C. The EV-containing supernatant is overlaid on a triple sucrose cushion (0.6 M, 1.3 M, 2.5 M) and ultracentrifuged for 3 h at 180,000 × g to separate vesicles based on density. The top of the gradient is discarded and fractions designated 1, 2 and 3 are collected and the refractive index is measured. Each fraction is further ultracentrifuged at 100,000 × g to pellet the vesicles contained in each fraction. Each preparation is validated by a combination of techniques including electron microscopy and RNA and protein analysis. Note – some tissue samples will not be amenable to this method. Post-mortem delay, storage time and the number of freeze-thaw cycles will negatively impact on tissue quality and result in contamination of the fractions with cellular debris and non-exosome vesicles.

Techniques Used: Isolation, Incubation, Electron Microscopy

Modification:

Article Title: SMAD3 Deficiency Promotes Inflammatory Aortic Aneurysms in Angiotensin II-Infused Mice Via Activation of iNOS
Article Snippet: .. Briefly, the whole aorta was dissected out from its origin at the left ventricle to the iliac bifurcation, denuded of periaortic fat, cut into small pieces (≈2‐mm segments), and incubated in 5 mL of Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (#16000036; Life Technologies) and 1.36 mg/mL of type II collagenase (#LS004174; Worthington Biochemical Corporation) in agitation at 115 rpm for 2 hours at 37°C. .. After digestion, 5 mL of medium was added, and the cells were centrifuged at 300g at 4°C for 5 minutes to pellet the cells for flow cytometric analysis.

Isolation:

Article Title: Exogenous Stimulation of Human Intervertebral Disc Cells in 3-Dimensional Alginate Bead Culture With BMP2 and L51P: Cytocompatibility and Effects on Cell Phenotype
Article Snippet: .. Briefly, cells were isolated by pronase (Roche, Basel, Switzerland) followed by collagenase type 2 (Worthington, London, UK) enzymatic digestion as previously reported. .. NPCs, AFCs and CEPCs were expanded in proliferation medium (low-glucose [1g/L] Dulbecco’s Modified Eagle Medium supplemented with 10% foetal bovine serum [FBS] and penicillin/streptomycin [P/S]).

Article Title: O-GlcNAc transferase inhibits visceral fat lipolysis and promotes diet-induced obesity
Article Snippet: .. For brown adipocyte isolation, freshly dissected interscapular BAT was cut into ~2 mm diameter pieces, digested in Krebs-Ringer buffer supplemented with 1% fatty acid free BSA, 0.1% HEPES (1 M, pH 7.3), 0.8 mg/mL Type 2 Collagenase (Worthington, LS004176), and 1.2 mM calcium chloride at 37 °C for 50 min. Digested tissue was then filtered through a 100 μm membrane and centrifuged at 300 g for 3 min. ..

Incubation:

Article Title: Fli1-haploinsufficient dermal fibroblasts promote skin-localized transdifferentiation of Th2-like regulatory T cells
Article Snippet: .. Dermis was minced and then incubated with 2 mg/ml collagenase type 2 (CLS-2; Worthington Biochemical, Lakewood, NJ, USA) in Tyrode’s solution for 60–90 minutes. ..

Mouse Assay:

Article Title: Cardiac O-GlcNAcylation blunts autophagic signaling in the diabetic heart
Article Snippet: .. Briefly, hearts were rapidly excised from heparinized and anesthetized mice, perfused retrogradely with Ca2+ -free perfusion buffer consisting of (in mM) 0.6 KH2 PO4 , 0.6 Na2 HPO4 , 1.2 MgSO4 • 7H2 O, 0.032 phenol red, 12 NaHCO3 , 10 KHCO3 , 10 HEPES, 30 taurine, 10 2,3-butanedione monoxime, and 5.5 glucose, pH 7.46, followed by buffer containing 12.5 μM CaCl2 and 0.4 mg/ml collagenase type 2 (Worthington). ..