pds kit inhibitor vial  (Worthington Biochemical)


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    Name:
    PDS Kit Inhibitor Vial
    Description:
    Ovomucoid protease inhibitor and bovine serum albumin which is 0 22 micron filtered and lyophilized in autoclaved vials to contain 10 mg ml each upon reconstitution with 32 ml of EBSS
    Catalog Number:
    lk003182
    Price:
    72
    Size:
    1 vi
    Source:
    Egg White/BSA
    Cas Number:
    9035.81.8/9048.46.8
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    Structured Review

    Worthington Biochemical pds kit inhibitor vial
    Ovomucoid protease inhibitor and bovine serum albumin which is 0 22 micron filtered and lyophilized in autoclaved vials to contain 10 mg ml each upon reconstitution with 32 ml of EBSS
    https://www.bioz.com/result/pds kit inhibitor vial/product/Worthington Biochemical
    Average 94 stars, based on 1521 article reviews
    Price from $9.99 to $1999.99
    pds kit inhibitor vial - by Bioz Stars, 2020-08
    94/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: Age-dependent accumulation of oligomeric SNCA/α-synuclein from impaired degradation in mutant LRRK2 knockin mouse model of Parkinson disease: role for therapeutic activation of chaperone-mediated autophagy (CMA)
    Article Snippet: .. Measurements of total cellular LAMP2A and GAPDH (CMA-substrate) levels in mouse ventral midbrain in flow cytometry Aged 18-month old WT and LRRK2R1441G KI mutant mouse ventral midbrains were freshly dissected and dissociated by papain dissociation system (Worthington Biochemical Corporation, LK003182) at 37°C with gentle agitation. .. Cells were passed through nylon mesh (70 micron) to obtain single cell suspension, and were fixed with 4% PFA (paraformaldehyde; Affymetrix Incorporation, 19943) at RT for 10 min.

    Filtration:

    Article Title: Single-cell transcriptomics of the aged mouse brain reveals convergent, divergent and unique aging signatures
    Article Snippet: .. Meninges were also removed, and the remaining brain tissue was dissociated into single cells using the Adult Brain Dissociation kit (Miltenyi Biotec #130-107-677) with these modifications: (a) the tissue was manually dissociated following the basic steps of the protocol described in the Neural Tissue Dissociation Kit (Miltenyi Biotec #130-092-628); (b) 5% (w/v) trehalose (Sigma Aldrich #T0167) was added in all buffers to ensure higher cellular viability ; (c) half concentration of papain was used, and the digestion was performed at 33-35°C; (d) the enzymatic reaction was quenched using ovomucoid protease inhibitor, as described in the Papain Dissociation System (Worthington #LK003182); (e) cell clusters were removed by serial filtration through pre-wetted 70um (Falcon #352350) and 40um (Falcon #352340) nylon cell strainers; (f) myelin debris and erythrocyte removal steps were omitted to prevent any bias in the recovered cell yields; (g) all centrifugations were performed at 220xg for 8min at 4°C. .. Single-cell RNA-sequencing For the scRNA-seq experiments, 8 young and 8 old mouse brains were analyzed, with 2 animals sacrificed per day.

    Concentration Assay:

    Article Title: Single-cell transcriptomics of the aged mouse brain reveals convergent, divergent and unique aging signatures
    Article Snippet: .. Meninges were also removed, and the remaining brain tissue was dissociated into single cells using the Adult Brain Dissociation kit (Miltenyi Biotec #130-107-677) with these modifications: (a) the tissue was manually dissociated following the basic steps of the protocol described in the Neural Tissue Dissociation Kit (Miltenyi Biotec #130-092-628); (b) 5% (w/v) trehalose (Sigma Aldrich #T0167) was added in all buffers to ensure higher cellular viability ; (c) half concentration of papain was used, and the digestion was performed at 33-35°C; (d) the enzymatic reaction was quenched using ovomucoid protease inhibitor, as described in the Papain Dissociation System (Worthington #LK003182); (e) cell clusters were removed by serial filtration through pre-wetted 70um (Falcon #352350) and 40um (Falcon #352340) nylon cell strainers; (f) myelin debris and erythrocyte removal steps were omitted to prevent any bias in the recovered cell yields; (g) all centrifugations were performed at 220xg for 8min at 4°C. .. Single-cell RNA-sequencing For the scRNA-seq experiments, 8 young and 8 old mouse brains were analyzed, with 2 animals sacrificed per day.

    Mutagenesis:

    Article Title: Age-dependent accumulation of oligomeric SNCA/α-synuclein from impaired degradation in mutant LRRK2 knockin mouse model of Parkinson disease: role for therapeutic activation of chaperone-mediated autophagy (CMA)
    Article Snippet: .. Measurements of total cellular LAMP2A and GAPDH (CMA-substrate) levels in mouse ventral midbrain in flow cytometry Aged 18-month old WT and LRRK2R1441G KI mutant mouse ventral midbrains were freshly dissected and dissociated by papain dissociation system (Worthington Biochemical Corporation, LK003182) at 37°C with gentle agitation. .. Cells were passed through nylon mesh (70 micron) to obtain single cell suspension, and were fixed with 4% PFA (paraformaldehyde; Affymetrix Incorporation, 19943) at RT for 10 min.

    Protease Inhibitor:

    Article Title: Dedifferentiation of committed epithelial cells into stem cells in vivo
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. # LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 minutes. .. Cells were then pelleted and stained with EpCAM-PECy7 (1:50; 25-5791-80, eBiosciences) or EpCAM-APC (1:50; 17-5791, eBiosciences); GSIβ4 (Griffonia Simplicifolia Isolectin beta4 )-Biotin (L2120, Sigma); SSEA1 eFluor 650NC (1:75, 95-8813-41, eBiosciences); PE anti-mouse CD24 (1:100, 553262, BD Pharmingen); for 30 min in 2.5% FBS in PBS on ice.

    Article Title: A revised airway epithelial hierarchy includes CFTR-expressing ionocytes
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. no. LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 min. .. Cells were then pelleted and stained with EpCAM–PECy7 (1:50; 25–5791-80, eBioscience) and CD45, CD81, or basis of GFP expression for 30 min in 2.5% FBS in PBS on ice.

    Article Title: Single-cell transcriptomics of the aged mouse brain reveals convergent, divergent and unique aging signatures
    Article Snippet: .. Meninges were also removed, and the remaining brain tissue was dissociated into single cells using the Adult Brain Dissociation kit (Miltenyi Biotec #130-107-677) with these modifications: (a) the tissue was manually dissociated following the basic steps of the protocol described in the Neural Tissue Dissociation Kit (Miltenyi Biotec #130-092-628); (b) 5% (w/v) trehalose (Sigma Aldrich #T0167) was added in all buffers to ensure higher cellular viability ; (c) half concentration of papain was used, and the digestion was performed at 33-35°C; (d) the enzymatic reaction was quenched using ovomucoid protease inhibitor, as described in the Papain Dissociation System (Worthington #LK003182); (e) cell clusters were removed by serial filtration through pre-wetted 70um (Falcon #352350) and 40um (Falcon #352340) nylon cell strainers; (f) myelin debris and erythrocyte removal steps were omitted to prevent any bias in the recovered cell yields; (g) all centrifugations were performed at 220xg for 8min at 4°C. .. Single-cell RNA-sequencing For the scRNA-seq experiments, 8 young and 8 old mouse brains were analyzed, with 2 animals sacrificed per day.

    Activation Assay:

    Article Title: Parent stem cells can serve as niches for their own daughter cells
    Article Snippet: .. Briefly, following trachea removal, airway tissue was cut into small fragments and transferred to a 2 ml solution containing 1ml 100 U of pre-activated papain (Worthington biochemical Corporation, cat. # LK003182) and 1 ml of activation buffer as per the manufacturer’s protocol. .. Tissue fragments were incubated on a shaking platform for 90 minutes at 37°C. the cell suspension was passed through a 70μm cell strainer to remove airway husks and pelleted for 5 minutes at 400g.

    Incubation:

    Article Title: Dedifferentiation of committed epithelial cells into stem cells in vivo
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. # LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 minutes. .. Cells were then pelleted and stained with EpCAM-PECy7 (1:50; 25-5791-80, eBiosciences) or EpCAM-APC (1:50; 17-5791, eBiosciences); GSIβ4 (Griffonia Simplicifolia Isolectin beta4 )-Biotin (L2120, Sigma); SSEA1 eFluor 650NC (1:75, 95-8813-41, eBiosciences); PE anti-mouse CD24 (1:100, 553262, BD Pharmingen); for 30 min in 2.5% FBS in PBS on ice.

    Article Title: A revised airway epithelial hierarchy includes CFTR-expressing ionocytes
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. no. LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 min. .. Cells were then pelleted and stained with EpCAM–PECy7 (1:50; 25–5791-80, eBioscience) and CD45, CD81, or basis of GFP expression for 30 min in 2.5% FBS in PBS on ice.

    Activity Assay:

    Article Title: Parent stem cells can serve as niches for their own daughter cells
    Article Snippet: .. The supernatant was aspirated and the pellet was resuspended in ovomucoid solution (Worthington biochemical Corporation, cat. # LK003182) for 20 minutes at 4°C to inactivate residual papain activity. .. Dissociated cells were stained with the following antibodies: EpCAM-PECy7 (1:50; 25-5791-80, eBiosciences) or EpCAM-APC (1:50; 17-5791, eBiosciences); GSIβ4 (Griffonia Simplicifolia Isolectin beta4)-Biotin (L2120, Sigma); SSEA-1 eFluor 650NC (1:75, 95-8813-41, eBiosciences); CD24-PE (1:100, 553262, BD Pharmingen).

    Article Title: Dedifferentiation of committed epithelial cells into stem cells in vivo
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. # LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 minutes. .. Cells were then pelleted and stained with EpCAM-PECy7 (1:50; 25-5791-80, eBiosciences) or EpCAM-APC (1:50; 17-5791, eBiosciences); GSIβ4 (Griffonia Simplicifolia Isolectin beta4 )-Biotin (L2120, Sigma); SSEA1 eFluor 650NC (1:75, 95-8813-41, eBiosciences); PE anti-mouse CD24 (1:100, 553262, BD Pharmingen); for 30 min in 2.5% FBS in PBS on ice.

    Article Title: A revised airway epithelial hierarchy includes CFTR-expressing ionocytes
    Article Snippet: .. Cell pellets were dispersed and incubated with Ovo-mucoid protease inhibitor (Worthington biochemical Corporation, cat. no. LK003182) to inactivate residual papain activity by incubating on a rocker at 4°C for 20 min. .. Cells were then pelleted and stained with EpCAM–PECy7 (1:50; 25–5791-80, eBioscience) and CD45, CD81, or basis of GFP expression for 30 min in 2.5% FBS in PBS on ice.

    Isolation:

    Article Title: Glial type specific regulation of CNS angiogenesis by HIFα-activated different signaling pathways
    Article Snippet: .. The isolated cortical tissues were dissociated by papain dissociation kit (#LK003176, Worthington) supplemented with DNase I (250 U/ml; #D5025, Sigma) and D-(+)-glucose (0.36%; #0188 AMRESCO) in 33 °C/10% CO2 for 90 min. Next, the tissues were transferred in PDS Kit-Inhibitor solution (#LK003182, Worthington). ..

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  • 94
    Worthington Biochemical pds kit inhibitor vial
    Pds Kit Inhibitor Vial, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pds kit inhibitor vial/product/Worthington Biochemical
    Average 94 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
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    92
    Worthington Biochemical pd l2
    Impact of <t>RGMb–PD-L2</t> blockade on tolerance induction is not caused by depletion of APC. (a–c) Mice immunized with OVA in ALUM on day 0 received OVA i.n. on days 7–9 and were treated with PD-L2 mAb 2C9 or isotype control (500 µg i.p.) on day 8. Lung cells were dispersed on day 10 and the expression of PD-L2 analyzed on AMs (F4/80 + CD11c + ), DCs (F4/80 − CD11c + ), and IMs (F4/80 + CD11c lo ). (a) Gating strategy used in b–d. (b) Solid line indicates staining with PD-L2 mAb TY25; shaded histogram indicates isotype control. One mouse/group representative of three is shown. (c) The number of PD-L2 + macrophages and DCs was quantified. Data are representative of two experiments.
    Pd L2, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pd l2/product/Worthington Biochemical
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    84
    Worthington Biochemical in vivo pd l1 expression
    PD-L1 expression increases in response to 4H-CTX or paclitaxel (PTX) treatment in vitro. a  EMT6-CDDP cells were treated with increasing concentrations of 4H-CTX, which resulted in increased expression of PD-L1 analyzed by flow cytometry.  b  EMT6-CDDP,  c  EMT6/P, or  d  SP1-AC2M2 cells were treated with 20 µM 4-HCTX or 1 µM of PTX; values represent mean fluorescence intensity of PD-L1 analyzed by flow cytometry with subtraction of mean fluorescence intensity of isotype IgG. One-way analysis of variance with Tukey’s multiple comparison test. Unpaired  t -test for SP1-AC2M2 analysis. * p
    In Vivo Pd L1 Expression, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    thermo fisher pds kit ebss vial
    PD-L1 expression increases in response to 4H-CTX or paclitaxel (PTX) treatment in vitro. a  EMT6-CDDP cells were treated with increasing concentrations of 4H-CTX, which resulted in increased expression of PD-L1 analyzed by flow cytometry.  b  EMT6-CDDP,  c  EMT6/P, or  d  SP1-AC2M2 cells were treated with 20 µM 4-HCTX or 1 µM of PTX; values represent mean fluorescence intensity of PD-L1 analyzed by flow cytometry with subtraction of mean fluorescence intensity of isotype IgG. One-way analysis of variance with Tukey’s multiple comparison test. Unpaired  t -test for SP1-AC2M2 analysis. * p
    Pds Kit Ebss Vial, supplied by thermo fisher, used in various techniques. Bioz Stars score: 80/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Impact of RGMb–PD-L2 blockade on tolerance induction is not caused by depletion of APC. (a–c) Mice immunized with OVA in ALUM on day 0 received OVA i.n. on days 7–9 and were treated with PD-L2 mAb 2C9 or isotype control (500 µg i.p.) on day 8. Lung cells were dispersed on day 10 and the expression of PD-L2 analyzed on AMs (F4/80 + CD11c + ), DCs (F4/80 − CD11c + ), and IMs (F4/80 + CD11c lo ). (a) Gating strategy used in b–d. (b) Solid line indicates staining with PD-L2 mAb TY25; shaded histogram indicates isotype control. One mouse/group representative of three is shown. (c) The number of PD-L2 + macrophages and DCs was quantified. Data are representative of two experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: Impact of RGMb–PD-L2 blockade on tolerance induction is not caused by depletion of APC. (a–c) Mice immunized with OVA in ALUM on day 0 received OVA i.n. on days 7–9 and were treated with PD-L2 mAb 2C9 or isotype control (500 µg i.p.) on day 8. Lung cells were dispersed on day 10 and the expression of PD-L2 analyzed on AMs (F4/80 + CD11c + ), DCs (F4/80 − CD11c + ), and IMs (F4/80 + CD11c lo ). (a) Gating strategy used in b–d. (b) Solid line indicates staining with PD-L2 mAb TY25; shaded histogram indicates isotype control. One mouse/group representative of three is shown. (c) The number of PD-L2 + macrophages and DCs was quantified. Data are representative of two experiments.

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Mouse Assay, Expressing, Affinity Magnetic Separation, Staining

    Blockade of PD-L2–RGMb interaction reduces the initial expansion of antigen-specific T cells after intranasal OVA administration. (a) Experimental protocol for Fig. 9 . Mice received DO11.10 T cells on day 0 and OVA i.n. The number of KJ1-26 + cells in mediastinal LNs was determined on the indicated days. (b) Mice treated with RGMb mAb 9D1 or control mAb on day −1 received 5 × 10 6 DO11.10 T cells. Left, TCRβ + KJ1-26 + CD4 + ; Right, FoxP3 + TCRβ + KJ1-26 + CD4 + cells. For each group, mediastinal LNs of 3–5 mice were combined; data are representative of two experiments. (c–e) Mice received 2 × 10 5 DO11.10 T cells after (c) RGMb mAb 9D1 or control mAb; (d) RGMb mAb 9D1, PD-L2 mAb 3.2, or control mAb; (e) PD-L2 mAb 3.2 or control mAb. (f) WT DO11.10 T cells were transferred into WT or PD-L2 −/− mice; (g) WT or PD-L2 −/− DO11.10 T cells were transferred into WT mice. (h–j) Mice received 2 × 10 5 DO11.10 T cells after (h) PD-L2 mAb 2C9 or control mAb; (i) PD-L1 mAb 9G2 or control mAb; (j) PD-L2 mAb 3.2, PD-1 mAb 1A12, or control mAb. The number of KJ1-26 + T cells in mediastinal lymph nodes on day 5 (h–j) was determined by flow cytometry. (b–d) Each point represents a pool of LNs from three mice. Data are representative of two to four experiments. (e–j) Each point represents the mean and SEM of KJ1-26 + T cells in LNs of two or three groups of two mice each. Data are representative of two experiments. **, P

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: Blockade of PD-L2–RGMb interaction reduces the initial expansion of antigen-specific T cells after intranasal OVA administration. (a) Experimental protocol for Fig. 9 . Mice received DO11.10 T cells on day 0 and OVA i.n. The number of KJ1-26 + cells in mediastinal LNs was determined on the indicated days. (b) Mice treated with RGMb mAb 9D1 or control mAb on day −1 received 5 × 10 6 DO11.10 T cells. Left, TCRβ + KJ1-26 + CD4 + ; Right, FoxP3 + TCRβ + KJ1-26 + CD4 + cells. For each group, mediastinal LNs of 3–5 mice were combined; data are representative of two experiments. (c–e) Mice received 2 × 10 5 DO11.10 T cells after (c) RGMb mAb 9D1 or control mAb; (d) RGMb mAb 9D1, PD-L2 mAb 3.2, or control mAb; (e) PD-L2 mAb 3.2 or control mAb. (f) WT DO11.10 T cells were transferred into WT or PD-L2 −/− mice; (g) WT or PD-L2 −/− DO11.10 T cells were transferred into WT mice. (h–j) Mice received 2 × 10 5 DO11.10 T cells after (h) PD-L2 mAb 2C9 or control mAb; (i) PD-L1 mAb 9G2 or control mAb; (j) PD-L2 mAb 3.2, PD-1 mAb 1A12, or control mAb. The number of KJ1-26 + T cells in mediastinal lymph nodes on day 5 (h–j) was determined by flow cytometry. (b–d) Each point represents a pool of LNs from three mice. Data are representative of two to four experiments. (e–j) Each point represents the mean and SEM of KJ1-26 + T cells in LNs of two or three groups of two mice each. Data are representative of two experiments. **, P

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    Blocking capacities of RGMb and PD-L2 mAbs and a model for RGMb and PD-L2 interactions. (a–c) The blocking capacity of RGMb mAbs and PD-L2 mAbs was determined by cell conjugation assay. After cells were stained with dyes, (a) 300-mRGMb cells were incubated with the indicated concentrations of RGMb mAbs before incubation with 300-mPD-L2 cells; (b) 300-mPD-L2 cells were incubated with the indicated concentrations of PD-L2 mAbs before incubation with 300-mRGMb cells; or (c) before incubation with 300-mPD-1 cells. (d) mRGMb-HIS was preincubated with mAbs or Ig fusion proteins and then added to BMP-4–coated plates. Binding of mRGMb-HIS was detected by anti–penta-HIS-HRP in an ELISA. Similar results were seen with BMP-2 (not depicted). (e) mPD-L2-hIgG or control-hIgG were preincubated alone or with monomeric mRGMb-HIS, and then added to BMP-4–coated plates. Binding of Ig fusion proteins was detected with anti–hIgG-HRP in an ELISA. Similar results were also seen for BMP-2 (not depicted). (f) mNeogenin- or mPD-L2–transfected 300 cells were stained with indicated Ig fusion proteins or control-Ig and analyzed by flow cytometry. (g) Binding of 300-mRGMb cells to 300-mPD-L2 or 300-mNeogenin cells was analyzed by cell conjugation assay. Cells stained with a red dye were incubated with cells stained with a green dye and binding was measured by flow cytometry. (h) Molecular model depicting PDL2–BMP–BMPR–RGMb–neogenin and PD-L2–PD-1 pathways. All data are representative of two or more independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: Blocking capacities of RGMb and PD-L2 mAbs and a model for RGMb and PD-L2 interactions. (a–c) The blocking capacity of RGMb mAbs and PD-L2 mAbs was determined by cell conjugation assay. After cells were stained with dyes, (a) 300-mRGMb cells were incubated with the indicated concentrations of RGMb mAbs before incubation with 300-mPD-L2 cells; (b) 300-mPD-L2 cells were incubated with the indicated concentrations of PD-L2 mAbs before incubation with 300-mRGMb cells; or (c) before incubation with 300-mPD-1 cells. (d) mRGMb-HIS was preincubated with mAbs or Ig fusion proteins and then added to BMP-4–coated plates. Binding of mRGMb-HIS was detected by anti–penta-HIS-HRP in an ELISA. Similar results were seen with BMP-2 (not depicted). (e) mPD-L2-hIgG or control-hIgG were preincubated alone or with monomeric mRGMb-HIS, and then added to BMP-4–coated plates. Binding of Ig fusion proteins was detected with anti–hIgG-HRP in an ELISA. Similar results were also seen for BMP-2 (not depicted). (f) mNeogenin- or mPD-L2–transfected 300 cells were stained with indicated Ig fusion proteins or control-Ig and analyzed by flow cytometry. (g) Binding of 300-mRGMb cells to 300-mPD-L2 or 300-mNeogenin cells was analyzed by cell conjugation assay. Cells stained with a red dye were incubated with cells stained with a green dye and binding was measured by flow cytometry. (h) Molecular model depicting PDL2–BMP–BMPR–RGMb–neogenin and PD-L2–PD-1 pathways. All data are representative of two or more independent experiments.

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Blocking Assay, Conjugation Assay, Staining, Incubation, Binding Assay, Enzyme-linked Immunosorbent Assay, Transfection, Flow Cytometry, Cytometry

    Expression of RGMb, PD-L2, BMPs, BMPRs, and related molecules in resting lung cell populations and airway epithelial cells. Expression of indicated mRNAs in resting lung cell populations and airway epithelial cells by qRT-PCR. (a–h) IMs (F4/80 + CD11c − ), AMs (F4/80 + CD11c + ), DCs (F4/80 − CD11c + ), and other cells (F4/80 − CD11c − ) were sorted from lung cells pooled from five to eight mice; CD4 + T cells (TCRβ + CD4 + ) and CD8 + T cells (TCRβ + CD8 + ) were sorted from lung cells pooled from two mice. (i) AECs and TECs were pooled from three and eight mice, respectively. Data are mean ± SEM; n = 3; *, P

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: Expression of RGMb, PD-L2, BMPs, BMPRs, and related molecules in resting lung cell populations and airway epithelial cells. Expression of indicated mRNAs in resting lung cell populations and airway epithelial cells by qRT-PCR. (a–h) IMs (F4/80 + CD11c − ), AMs (F4/80 + CD11c + ), DCs (F4/80 − CD11c + ), and other cells (F4/80 − CD11c − ) were sorted from lung cells pooled from five to eight mice; CD4 + T cells (TCRβ + CD4 + ) and CD8 + T cells (TCRβ + CD8 + ) were sorted from lung cells pooled from two mice. (i) AECs and TECs were pooled from three and eight mice, respectively. Data are mean ± SEM; n = 3; *, P

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Expressing, Quantitative RT-PCR, Affinity Magnetic Separation, Mouse Assay

    RGMb binds to PD-L2, but not PD-L1 or other related molecules. (a and b) mRGMb- or mPD-L2–transfected 300 cells, untransfected 300 cells, and PD-1–transfected Jurkat T cells were stained with the indicated Ig fusion proteins (red) or control-Ig (blue) and analyzed by flow cytometry. MFI, mean fluorescence intensity. (c and d) Binding of mPD-L2-Ig or control-Ig to plates coated with mRGMb-HIS, hRGMb-HIS, mRGMa-HIS or mRGMc-HIS, was analyzed by ELISA. (e) Biacore analysis of the interaction of RGMb with PD-L2 and of PD-1 with PD-L2. (f–k) Cell–cell binding of the indicated transfected cells was analyzed by cell conjugation assay. In f, h, and j, the top panels show the FSC-SSC plots and the bottom panels show the corresponding dot plots. g, i, and k show only the dot plots. All data are representative of 2–11 independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: RGMb binds to PD-L2, but not PD-L1 or other related molecules. (a and b) mRGMb- or mPD-L2–transfected 300 cells, untransfected 300 cells, and PD-1–transfected Jurkat T cells were stained with the indicated Ig fusion proteins (red) or control-Ig (blue) and analyzed by flow cytometry. MFI, mean fluorescence intensity. (c and d) Binding of mPD-L2-Ig or control-Ig to plates coated with mRGMb-HIS, hRGMb-HIS, mRGMa-HIS or mRGMc-HIS, was analyzed by ELISA. (e) Biacore analysis of the interaction of RGMb with PD-L2 and of PD-1 with PD-L2. (f–k) Cell–cell binding of the indicated transfected cells was analyzed by cell conjugation assay. In f, h, and j, the top panels show the FSC-SSC plots and the bottom panels show the corresponding dot plots. g, i, and k show only the dot plots. All data are representative of 2–11 independent experiments.

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Transfection, Staining, Flow Cytometry, Cytometry, Fluorescence, Binding Assay, Enzyme-linked Immunosorbent Assay, Conjugation Assay

    PD-L2 or RGMb blockade inhibits induction of respiratory tolerance. (a) Experimental protocol for induction of respiratory tolerance. (b–d) PD-L2–deficient or WT mice in b, and WT mice injected with the indicated mAb in c and d, were exposed to OVA i.n. (tolerized) or PBS (nontolerized) and subsequently received 50 µg OVA/ALUM i.p. T cell proliferation (top) and IL-4 production (bottom) in response to restimulation with OVA in vitro are shown. n = 2–3. Data are representative of two to five experiments. Data are mean ± SEM, *, P

    Journal: The Journal of Experimental Medicine

    Article Title: RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance

    doi: 10.1084/jem.20130790

    Figure Lengend Snippet: PD-L2 or RGMb blockade inhibits induction of respiratory tolerance. (a) Experimental protocol for induction of respiratory tolerance. (b–d) PD-L2–deficient or WT mice in b, and WT mice injected with the indicated mAb in c and d, were exposed to OVA i.n. (tolerized) or PBS (nontolerized) and subsequently received 50 µg OVA/ALUM i.p. T cell proliferation (top) and IL-4 production (bottom) in response to restimulation with OVA in vitro are shown. n = 2–3. Data are representative of two to five experiments. Data are mean ± SEM, *, P

    Article Snippet: To induce tolerance, lightly anesthetized WT BALB/cByJ or PD-L2−/− mice received 100 µg of LPS-free OVA (Worthington Biochemical) in PBS or PBS alone (control) by intranasal instillation on days 0, 1, and 2.

    Techniques: Mouse Assay, Injection, In Vitro

    PD-L1 expression increases in response to 4H-CTX or paclitaxel (PTX) treatment in vitro. a  EMT6-CDDP cells were treated with increasing concentrations of 4H-CTX, which resulted in increased expression of PD-L1 analyzed by flow cytometry.  b  EMT6-CDDP,  c  EMT6/P, or  d  SP1-AC2M2 cells were treated with 20 µM 4-HCTX or 1 µM of PTX; values represent mean fluorescence intensity of PD-L1 analyzed by flow cytometry with subtraction of mean fluorescence intensity of isotype IgG. One-way analysis of variance with Tukey’s multiple comparison test. Unpaired  t -test for SP1-AC2M2 analysis. * p

    Journal: NPJ Breast Cancer

    Article Title: Immunostimulatory and anti-tumor metronomic cyclophosphamide regimens assessed in primary orthotopic and metastatic murine breast cancer

    doi: 10.1038/s41523-020-0171-1

    Figure Lengend Snippet: PD-L1 expression increases in response to 4H-CTX or paclitaxel (PTX) treatment in vitro. a EMT6-CDDP cells were treated with increasing concentrations of 4H-CTX, which resulted in increased expression of PD-L1 analyzed by flow cytometry. b EMT6-CDDP, c EMT6/P, or d SP1-AC2M2 cells were treated with 20 µM 4-HCTX or 1 µM of PTX; values represent mean fluorescence intensity of PD-L1 analyzed by flow cytometry with subtraction of mean fluorescence intensity of isotype IgG. One-way analysis of variance with Tukey’s multiple comparison test. Unpaired t -test for SP1-AC2M2 analysis. * p

    Article Snippet: For analysis of in vivo PD-L1 expression, tumors were dissociated with the following enzymatic buffer (1% BSA, 12,500 units collagenase II (Worthington), 12,500 units collagenase IV (Worthington), and DNase I (cat. no. LS006333, Worthington) for 45 min and then a single-cell suspension gained.

    Techniques: Expressing, In Vitro, Flow Cytometry, Fluorescence