scid beige mice  (Worthington Biochemical)


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    Structured Review

    Worthington Biochemical scid beige mice
    Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized <t>SCID/beige</t> mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to <t>Ki67</t> IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies
    Scid Beige Mice, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 93/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Aberrant FGFR signaling mediates resistance to CDK4/6 inhibitors in ER+ breast cancer"

    Article Title: Aberrant FGFR signaling mediates resistance to CDK4/6 inhibitors in ER+ breast cancer

    Journal: Nature Communications

    doi: 10.1038/s41467-019-09068-2

    Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized SCID/beige mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to Ki67 IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies
    Figure Legend Snippet: Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized SCID/beige mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to Ki67 IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies

    Techniques Used: Inhibition, Amplification, Mouse Assay, Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Expressing, Staining

    Related Articles

    Isolation:

    Article Title: Collagen IV and basement membrane at the evolutionary dawn of metazoan tissues
    Article Snippet: .. Isolation, purification, and analysis of collagen IV NC1 hexamers Whole ctenophore tissues were frozen in liquid nitrogen, pulverized in a mortar and pestle and then homogenized in 2.0 ml g−1 digestion buffer and 0.1 mg ml−1 Worthington Biochemical bacterial collagenase and allowed to digest at 37°C, with spinning for 24 hr. .. Liquid chromatography purification of solubilized NC1 varied by species based on protein yield.

    Article Title: Extracellular chloride signals collagen IV network assembly during basement membrane formation
    Article Snippet: .. Isolation of NC1 domains from collagen IV matrix was accomplished by treating the matrix with bacterial collagenase enzyme (Worthington Biochemical Corporation) at 37°C in 10 mM Tris-HCl, pH 7.5, 10 mM CaCl2, 10 mM KI, 0.4 mM PMSF, 1 µg/ml aprotinin, and 1 µg/ml leupeptin. ..

    Blocking Assay:

    Article Title: The Short and Long Forms of Type XVIII Collagen Show Clear Tissue Specificities in Their Expression and Location in Basement Membrane Zones in Humans
    Article Snippet: .. To reduce the possibility of the epitopes recognized by type XVIII collagen-specific antibodies being masked in tissues, some of the sections were pretreated with 6 mol/L urea, 0.1 mol/L glycine, pH 3.5, for 1 hour at 4°C, with 10 U/ml bacterial collagenase (Worthington Biochemical Corp.) for 2 hours at 37°C, with 5% hyaluronidase (Sigma Chemical Co.) for 15 minutes at 37°C, with 0.2 U chondroitinase ABC (Boehringer Mannheim) at 37°C overnight, with 0.2 U N -glycosidase (Boehringer Mannheim) at 37°C for overnight, or with 0.7 U heparin lyase II and III (Sigma Chemical Co.) at 37°C for overnight before blocking by incubation with 3% bovine serum albumin in PBS. .. The construct QH48, corresponding to all type XVIII variants, and QH67, corresponding to the long variant, were expressed as DHFR fusion proteins in E. coli (Figure 1) , and the purified recombinant proteins were used to immunize rabbits.

    Purification:

    Article Title: Collagen IV and basement membrane at the evolutionary dawn of metazoan tissues
    Article Snippet: .. Isolation, purification, and analysis of collagen IV NC1 hexamers Whole ctenophore tissues were frozen in liquid nitrogen, pulverized in a mortar and pestle and then homogenized in 2.0 ml g−1 digestion buffer and 0.1 mg ml−1 Worthington Biochemical bacterial collagenase and allowed to digest at 37°C, with spinning for 24 hr. .. Liquid chromatography purification of solubilized NC1 varied by species based on protein yield.

    Incubation:

    Article Title: Matrix assembly, regulation, and survival functions of laminin and its receptors in embryonic stem cell differentiation
    Article Snippet: .. Duplicates of type IV collagen antibody immunoprecipitates were incubated with 5 U bacterial collagenase (CLSPA; Worthington Biochemical Corporation) at 37°C for 1 h. After collagenase digestion, the immunoprecipitates were washed twice in PBS and analyzed. .. Semi-quantitative RT-PCR Total RNA was isolated with TRIzol® reagent (Life Technologies) and reverse-transcribed to cDNA using SuperScript™ II reverse transcriptase (Life Technologies).

    Article Title: The Short and Long Forms of Type XVIII Collagen Show Clear Tissue Specificities in Their Expression and Location in Basement Membrane Zones in Humans
    Article Snippet: .. To reduce the possibility of the epitopes recognized by type XVIII collagen-specific antibodies being masked in tissues, some of the sections were pretreated with 6 mol/L urea, 0.1 mol/L glycine, pH 3.5, for 1 hour at 4°C, with 10 U/ml bacterial collagenase (Worthington Biochemical Corp.) for 2 hours at 37°C, with 5% hyaluronidase (Sigma Chemical Co.) for 15 minutes at 37°C, with 0.2 U chondroitinase ABC (Boehringer Mannheim) at 37°C overnight, with 0.2 U N -glycosidase (Boehringer Mannheim) at 37°C for overnight, or with 0.7 U heparin lyase II and III (Sigma Chemical Co.) at 37°C for overnight before blocking by incubation with 3% bovine serum albumin in PBS. .. The construct QH48, corresponding to all type XVIII variants, and QH67, corresponding to the long variant, were expressed as DHFR fusion proteins in E. coli (Figure 1) , and the purified recombinant proteins were used to immunize rabbits.

    Activity Assay:

    Article Title: Comparison of the enzymatic efficiency of Liberase TM and tumor dissociation enzyme: effect on the viability of cells digested from fresh and cryopreserved human ovarian cortex
    Article Snippet: .. Their activity is directed to break the peptide bonds in collagen (Collagenase), fibronectin, collagen IV, and to a lower extent collagen I, however, it does not cleave collagen V or laminin of neutral protease (V Worthington Biochemical Corporation). ..

    Cell Culture:

    Article Title: Characterization of intercellular communication and mitochondrial donation by mesenchymal stromal cells derived from the human lung
    Article Snippet: .. Cells were cultured in 24-well sized plates pre-coated with a solution of 0.1 mg/ml fibronectin (Sigma Aldrich), 0.03 mg/ml bovine collagen type 1 (Worthington) and 0.01 mg/ml bovine serum albumin (Bovogen). ..

    SDS Page:

    Article Title: Improved catalytic efficiency, thermophilicity, anti-salt and detergent tolerance of keratinase KerSMD by partially truncation of PPC domain
    Article Snippet: .. The SDS-PAGE analysis related to the degradation of insoluble type I collagen fiber (bovine achilles tendon, Worthington Biochemical Co., USA) was conducted. .. The collagen (1 mg) was treated with different enzymes (50 U keratinolytic activity) at 37 °C for 12 h in 50 mM glycine-NaOH buffer (pH 9.0).

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    Worthington Biochemical collagenase type iii
    Collagenase Type Iii, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Worthington Biochemical scid beige mice
    Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized <t>SCID/beige</t> mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to <t>Ki67</t> IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies
    Scid Beige Mice, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Worthington Biochemical klareskog l
    Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized <t>SCID/beige</t> mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to <t>Ki67</t> IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies
    Klareskog L, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized SCID/beige mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to Ki67 IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies

    Journal: Nature Communications

    Article Title: Aberrant FGFR signaling mediates resistance to CDK4/6 inhibitors in ER+ breast cancer

    doi: 10.1038/s41467-019-09068-2

    Figure Lengend Snippet: Combined inhibition of ER, CDK4/6, and FGFR1 inhibits the growth of ERα/ FGFR1 -amplified breast cancer PDXs. a ER+/HER2−/ FGFR1 -amplified TM00386 PDX fragments were established in ovariectomized SCID/beige mice supplemented with a 21-day release, 0.25-mg 17β-estradiol pellet. Once tumors reached ≥200 mm 3 , mice were randomized to the indicated treatment arms. Each data point represents the fold change in volume ± SEM ( n = 8 per arm; ANOVA test). b Bar graph showing the % change in volume in individual xenografts after 3 weeks of treatment relative to individual tumor volumes on day 0. c Xenografts were harvested after 1 week of treatment and FFPE tumor sections were subjected to Ki67 IHC as described in Methods. The percent of Ki67+ tumor cells (inset) was assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm. d TM00386 PDXs in mice treated with vehicle or fulvestrant plus palbociclib were harvested and snap frozen at the end of treatment. RNA was extracted and subjected to nanoString analysis as described in Methods. Heat map represents different gene expression levels between controls ( n = 4) and tumors treated with fulvestrant plus palbociclib ( n = 4). e , f TM00386 PDXs were harvested at the end of treatment. FFPE sections from the PDXs were subjected to IHC with p-RB ( e ) and CCND1 ( f ) antibodies as described in Methods. The percent of p-RB and CCND1-positive tumor cells and their staining intensity were assessed by an expert breast pathologist (P.G.E.) blinded to the treatment arm to generate an H -score (shown in Supplementary figures ). g TM00386 tumors were harvested at the end of treatment, 4 h after the last dose of palbociclib and erdafitinib and 24 h after the last dose of fulvestrant. Tumor lysates were prepared and subjected to immunoblot analyses with the indicated antibodies

    Article Snippet: Ki67 detection and cell sorting TM00386 PDXs established in ovariectomized SCID/beige mice were harvested and dissociated using digestion buffer [125 μg/mL DnaseI (#LS002138, Worthington), 10 μg/mL Dispase (#LS02109, Worthington), 500 μg/mL Collagenase 3 (#LS004182, Worthington), and 5× triple antibiotics (#15240-062, Invitrogen)] to generate single cells.

    Techniques: Inhibition, Amplification, Mouse Assay, Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Expressing, Staining