dispase  (Worthington Biochemical)


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    Name:
    Neutral Protease Dispase Purified
    Description:
    Animal Origin Free Chromatographically purified A lyophilized powder
    Catalog Number:
    LS02100
    Price:
    70
    Source:
    Bacillus polymyxa
    Size:
    10 mg
    Cas Number:
    42613.33.2
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    Structured Review

    Worthington Biochemical dispase
    <t>Dispase</t> treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected
    Animal Origin Free Chromatographically purified A lyophilized powder
    https://www.bioz.com/result/dispase/product/Worthington Biochemical
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dispase - by Bioz Stars, 2021-04
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    Images

    1) Product Images from "Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment"

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment

    Journal: Journal of Virology

    doi: 10.1128/JVI.02889-14

    Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected
    Figure Legend Snippet: Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected

    Techniques Used: Purification

    Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa
    Figure Legend Snippet: Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa

    Techniques Used: Binding Assay, Purification

    The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.
    Figure Legend Snippet: The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.

    Techniques Used: Mutagenesis, Binding Assay

    Related Articles

    Incubation:

    Article Title: Comparison of quinazoline and benzoylpyrazoline chemotypes targeting the CaVα-β interaction as antagonists of the N-type CaV2.2 channel
    Article Snippet: DRG neurons were isolated from ~100 g Sprague-Dawley female rats. .. DRG were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat# 11,965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, CatLS02104; Worthington, Lakewood, NJ) and collagenase type I (5 mg/mL, Cat# LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37°C under gentile agitation. .. Dissociated DRG neurons were then gently centrifuged to collect cells and resuspended with DRG media DMEM containing 1% penicillin/streptomycin sulfate, 30 ng/mL nerve growth factor, and 10% fetal bovine serum (Hyclone) before plating onto poly-D-lysine – and laminin-coated glass 12-mm coverslips.

    Article Title: A Transcriptomic Analysis of Differential Gene Expression during Chick Periocular Neural Crest Differentiation into Corneal Cells
    Article Snippet: .. All tissues were incubated in dispase (1.5 mg/ml; Worthington) at 38°C for 5 minutes, then rinsed twice in Ringer’s solution. ..

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment
    Article Snippet: Briefly, immunoisolated hexon was incubated for 30 min in Tris-maleate buffer (50 mM Trizma-maleate, 10 mM NaCl, 1 mM EDTA, and 0.1% Tween 20 [pH 4.4]), resuspended in the same buffer at pH 7.4 for the indicated amounts of time, and then exposed to purified rat brain cytoplasmic dynein at 4°C for 1.5 h. Hexon-bound dynein was assayed by immunoblotting. .. For controlled proteolysis, hexon or AdV capsids were incubated with a 6-fold molar excess of dispase (neutral protease; Worthington Biochemicals, Freehold, NJ) over hexon in dispase buffer (100 mM Trizma-maleate, 120 mM NaCl, 2.5 mM CaCl2 ), and the pH was adjusted to either pH 7.4 (neutral) or pH 4.4 (acidic) for 5 min at 37°C until the reaction was stopped by the addition of either protein sample buffer (PSB) or 5 mM EDTA to the mixture. .. Sedimentation analysis of hexon was performed on linear 5 to 20% sucrose gradients of a volume of 1.4 ml in Tris-maleate buffer (50 mM Trizma-maleate, 10 mM NaCl, 1 mM EDTA, and 0.1% Tween 20) at the indicated pH values.

    Article Title: High-resolution transcriptional dissection of in vivo Atoh1-mediated hair cell conversion in mature cochleae identifies Isl1 as a co-reprogramming factor
    Article Snippet: The Cq values were converted to expression levels using the equation Log2 (Ex) = (LOD-Ct) value—Cq. .. Cochleae were dissected out and incubated in solution (50% accutase (Innovative Cell Technologies, San Diego), 0.02% Trypsin(Thermofisher), 125 μg/ml Thermolysin (SIGMA-ALDRICH)) at 37°C for 3 min. 0.02 mg/ml collagenase IV (SIGMA-ALDRICH) and dispase (Worthington Biochemicals Corp) were further added to the enzyme solution and incubated for 4 min at 37°C. ..

    Isolation:

    Article Title: PDGFRα signalling promotes fibrogenic responses in collagen‐producing cells in Duchenne muscular dystrophy
    Article Snippet: .. Cells for molecular analysis were isolated by FACS from single cell preparations prepared by digesting a collection of limb muscles (TA, gastrocnemius, and quadriceps) with collagenase and dispase (Worthington Biochemical Corp, Lakewood, NJ, USA) , . .. RNA was isolated from cells or tissue as described previously .

    FACS:

    Article Title: PDGFRα signalling promotes fibrogenic responses in collagen‐producing cells in Duchenne muscular dystrophy
    Article Snippet: .. Cells for molecular analysis were isolated by FACS from single cell preparations prepared by digesting a collection of limb muscles (TA, gastrocnemius, and quadriceps) with collagenase and dispase (Worthington Biochemical Corp, Lakewood, NJ, USA) , . .. RNA was isolated from cells or tissue as described previously .

    Modification:

    Article Title: Cell Sheet Comprised of Mesenchymal Stromal Cells Overexpressing Stem Cell Factor Promotes Epicardium Activation and Heart Function Improvement in a Rat Model of Myocardium Infarction
    Article Snippet: Rat MSC were isolated from subcutaneous adipose tissue. .. Briefly, the tissue sample was fragmented under sterile conditions to a homogeneous mass and subjected to enzymatic treatment in Dulbecco’s modified Eagle’s medium (DMEM) medium (Gibco, Waltham, MA, USA) with 10 U/mL dispase (Worthington Biochemical, Lakewood, NJ, USA) and 200 U/mL type I collagenase (Worthington Biochemical, Lakewood, NJ, USA) at 37 °C for 30 min. After centrifugation for 10 min at 200× g the cell pellet was resuspended in DMEM and filtered through 40 μm nylon membrane. .. Isolated cells were maintained in 4.5 g/L D-glucose DMEM with 10% fetal bovine serum (FBS, Gibco, Waltham, MA, USA) and 1% penicillin/streptomycin solution at 37 °C and 5% CO2.

    Centrifugation:

    Article Title: Cell Sheet Comprised of Mesenchymal Stromal Cells Overexpressing Stem Cell Factor Promotes Epicardium Activation and Heart Function Improvement in a Rat Model of Myocardium Infarction
    Article Snippet: Rat MSC were isolated from subcutaneous adipose tissue. .. Briefly, the tissue sample was fragmented under sterile conditions to a homogeneous mass and subjected to enzymatic treatment in Dulbecco’s modified Eagle’s medium (DMEM) medium (Gibco, Waltham, MA, USA) with 10 U/mL dispase (Worthington Biochemical, Lakewood, NJ, USA) and 200 U/mL type I collagenase (Worthington Biochemical, Lakewood, NJ, USA) at 37 °C for 30 min. After centrifugation for 10 min at 200× g the cell pellet was resuspended in DMEM and filtered through 40 μm nylon membrane. .. Isolated cells were maintained in 4.5 g/L D-glucose DMEM with 10% fetal bovine serum (FBS, Gibco, Waltham, MA, USA) and 1% penicillin/streptomycin solution at 37 °C and 5% CO2.

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  • 99
    Worthington Biochemical collagenase type 2
    Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and <t>type</t> 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (
    Collagenase Type 2, 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
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    99
    Worthington Biochemical collagenase i
    [A] Digestion of BAT with <t>collagenase</t> I produces an adipose fraction harboring brown adipocytes and a stromal fraction composed primarily of endothelium separated by a fluid portion. [B] Deep-red fluorescence intensity images of centrifugation tubes containing
    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
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    99
    Worthington Biochemical collagenase type iii
    Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow <t>cytometry.</t> Tumours were analysed in <t>three</t> independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P
    Collagenase Type Iii, 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
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    Image Search Results


    Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and type 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: SMAD3 Deficiency Promotes Inflammatory Aortic Aneurysms in Angiotensin II-Infused Mice Via Activation of iNOS

    doi: 10.1161/JAHA.113.000269

    Figure Lengend Snippet: Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and type 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (

    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.

    Techniques: Concentration Assay, Construct, Incubation, Derivative Assay, Generated, Mouse Assay, Expressing, Ex Vivo

    [A] Digestion of BAT with collagenase I produces an adipose fraction harboring brown adipocytes and a stromal fraction composed primarily of endothelium separated by a fluid portion. [B] Deep-red fluorescence intensity images of centrifugation tubes containing

    Journal: Journal of materials chemistry. B, Materials for biology and medicine

    Article Title: Fluorescence Imaging of Interscapular Brown Adipose Tissue in Living Mice †

    doi: 10.1039/C4TB01914H

    Figure Lengend Snippet: [A] Digestion of BAT with collagenase I produces an adipose fraction harboring brown adipocytes and a stromal fraction composed primarily of endothelium separated by a fluid portion. [B] Deep-red fluorescence intensity images of centrifugation tubes containing

    Article Snippet: The minced tissue was digested with 2 mg/ml Collagenase I (Worthington Biochemical Corporation) in Hanks' Balanced Salt Solution (HBSS) (1.3 mM CaCl2 , 0.5 mM MgCl2 •6H2 O, 0.5 mM MgSO4 •7H2 O, 5.3 mM KCl, 0.40 mM KH2 PO4 , 4.2 mM NaHCO3 , 140 mM, NaCl, 0.3 mM Na2 HPO4 , pH 7.5) at 37 °C for 2 hours.

    Techniques: Fluorescence, Centrifugation

    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

    Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow cytometry. Tumours were analysed in three independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P

    Journal: Nature Communications

    Article Title: Targeting VEGF-A in myeloid cells enhances natural killer cell responses to chemotherapy and ameliorates cachexia

    doi: 10.1038/ncomms12528

    Figure Lengend Snippet: Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow cytometry. Tumours were analysed in three independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P

    Article Snippet: Flow cytometry analysis Tumours were digested with 0.1% collagenase type III (Worthington) and 1 U ml−1 DNAse I (Promega) and incubated at 37 °C for 1 h. One hundred and six tumour cells were incubated with Fc-Block (BD Biosciences, #553141 at 1/100) before labelling with fluorochrome-conjugated antibodies.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Activity Assay, Recombinant