rat monoclonal antibodies  (Thermo Fisher)


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  • 96
    Name:
    Rat anti Mouse IgD Secondary Antibody
    Description:
    Rat anti Mouse IgD Secondary Antibody for Flow
    Catalog Number:
    MIGD01
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Cell Analysis|Flow Cytometry Antibodies & Secondary Detection|Flow Cytometry Antibodies for CD Markers|Flow Cytometry
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    Structured Review

    Thermo Fisher rat monoclonal antibodies
    Rat anti Mouse IgD Secondary Antibody for Flow
    https://www.bioz.com/result/rat monoclonal antibodies/product/Thermo Fisher
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rat monoclonal antibodies - by Bioz Stars, 2021-06
    96/100 stars

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    Related Articles

    Western Blot:

    Article Title: Cholesterol Accumulation in Dendritic Cells Links the Inflammasome to Acquired Immunity
    Article Snippet: Cells were cultured in 96 well plates in DMEM supplemented with 10% FBS and 1% pen-strep, and incubated with or without 20 ng/ml GM-CSF for 24 h (CD11c+ ) or with and without 50 µg/ml rHDL (CSL-111, a kind gift from Dr. Samuel Wright, CSL Australia) (CD11b+ ) for 24 h. Levels of IL-1β, IL-18, IL-23, IL-12p70, and IL-6 were measured in the media using ELISA (R & D systems), and corrected for cell protein. .. For Western blot to assess caspase-1 cleavage in CD11b+ cells, a primary rat anti-mouse caspase-1 antibody (eBioscience) and a rat secondary antibody (Cell-Signaling) were used. .. For co-incubation experiments, CD11c+ DCs were incubated with T-cells isolated from 8 week old wild-type mice using CD3ε+ beads (Miltenyi Biotec) in a ratio 1:5 for 5 days.

    Mouse Assay:

    Article Title: A Toll-like receptor 2 agonist-fused antigen enhanced antitumor immunity by increasing antigen presentation and the CD8 memory T cells population
    Article Snippet: Depleting subpopulations of CD4+ or CD8+ T lymphocytes in mice The experimental protocols of T cell depletion were modified from a previous study [ ]. .. Briefly, groups of mice were treated intraperitoneally i.p. by injection with 0.5 mg of the rat anti-mouse CD4 antibody (clone GK1.5, eBioscience) and rat anti-mouse CD8 antibody (clone 53–6.72, eBioscience) to deplete CD4+ or CD8+ T lymphocytes, respectively. .. A total of 0.5 mg of rat IgG (Invitrogen) was used as a control antibody in experiments.

    Article Title: Sleep disruption impairs hematopoietic stem cell transplantation in mice
    Article Snippet: Serum corticosterone levels were determined using a standard ELISA assay for corticosterone (Millipore; Billerica, MA), as per the standard procedure recommended by the manufacture. .. Flow cytometry Mice were euthanized, using CO2, and bone marrow was harvested into PBS containing 2% fetal calf serum (FCS). .. Lineage staining was performed with fluorochrome-conjugated monoclonal antibodies (eBioscience) against Ter119 (15-5921), Gr1 (15-5931), Mac1 (15-0112), B220 (15-0452), CD3 (15-0031), CD4 (15-0041), and CD8 (15-0081) (all in 1:200 dilution).

    Article Title: Induction of metabolic quiescence defines the transitional to follicular B cell switch
    Article Snippet: For sorting, B cells were run on a SORP Aria II (BD Biosciences) and collected in RMPI media supplemented with 20% FBS. .. Mouse Flow Cytometry and Cell SortingEight-week-old C57BL/6 mice were used for analysis. ..

    Injection:

    Article Title: A Toll-like receptor 2 agonist-fused antigen enhanced antitumor immunity by increasing antigen presentation and the CD8 memory T cells population
    Article Snippet: Depleting subpopulations of CD4+ or CD8+ T lymphocytes in mice The experimental protocols of T cell depletion were modified from a previous study [ ]. .. Briefly, groups of mice were treated intraperitoneally i.p. by injection with 0.5 mg of the rat anti-mouse CD4 antibody (clone GK1.5, eBioscience) and rat anti-mouse CD8 antibody (clone 53–6.72, eBioscience) to deplete CD4+ or CD8+ T lymphocytes, respectively. .. A total of 0.5 mg of rat IgG (Invitrogen) was used as a control antibody in experiments.

    Flow Cytometry:

    Article Title: Sleep disruption impairs hematopoietic stem cell transplantation in mice
    Article Snippet: Serum corticosterone levels were determined using a standard ELISA assay for corticosterone (Millipore; Billerica, MA), as per the standard procedure recommended by the manufacture. .. Flow cytometry Mice were euthanized, using CO2, and bone marrow was harvested into PBS containing 2% fetal calf serum (FCS). .. Lineage staining was performed with fluorochrome-conjugated monoclonal antibodies (eBioscience) against Ter119 (15-5921), Gr1 (15-5931), Mac1 (15-0112), B220 (15-0452), CD3 (15-0031), CD4 (15-0041), and CD8 (15-0081) (all in 1:200 dilution).

    Article Title: Induction of metabolic quiescence defines the transitional to follicular B cell switch
    Article Snippet: For sorting, B cells were run on a SORP Aria II (BD Biosciences) and collected in RMPI media supplemented with 20% FBS. .. Mouse Flow Cytometry and Cell SortingEight-week-old C57BL/6 mice were used for analysis. ..

    Incubation:

    Article Title: Bone marrow stromal antigen 2 expressed in cancer cells promotes mammary tumor growth and metastasis
    Article Snippet: .. Cells were incubated with BrdU label (1:2000) for 20 hours, treated with a fixative/denaturing solution (30 minutes) and incubated with an anti-BrdU antibody (1:1000) and rat anti-mouse BST-2 antibody (1:200, eBioscience) for 1 hour at room temperature. .. Cells were washed and incubated with Alexa Fluor 594 anti-rat (Invitrogen, Waltham, MA, USA) and Alexa Fluor 488 anti-mouse (Invitrogen) secondary antibodies for 30 minutes at room temperature.

    Immunocytochemistry:

    Article Title: Time-Dependent Disruption of Oviduct Pacemaker Cells by Chlamydia Infection in Mice 1
    Article Snippet: Tissues were then blocked for 1 h at room temperature with bovine serum albumin (1% w/v; Sigma-Aldrich, St. Louis, MO) before being placed in primary antibody for 48 h at 4°C. .. To identify ICC-OVI, rat anti-mouse KIT monoclonal (ACK2, 5 μg ml−1 ; eBioscience Inc., San Diego, CA [ , ]) or goat anti-mouse stem cell factor receptor monoclonal (2 μg ml−1 ; R & D Systems Inc., Minneapolis, MN) antibodies were used. .. Cells expressing NOS2 and PTGS2 were identified with a rabbit anti-mouse NOS2 polyclonal (5 μg ml−1 ; BD Biosciences, San Jose, CA) and a rabbit anti-mouse PTGS2 polyclonal (5 μg ml−1 ; Enzo Life Sciences International, Inc., Plymouth Meeting, PA) respectively.

    Immunohistochemistry:

    Article Title: Gene Expression Profiling of Lacrimal Glands Identifies the Ectopic Expression of MHC II on Glandular Cells as a Presymptomatic Feature in a Mouse Model of Primary Sjögren's Syndrome
    Article Snippet: Indirect Immunofluorescence staining was performed for the analysis of CD3+ T cell and CD19+ B cell by using rat-anti-mouse CD3 (clone: 17A2, Biolegend) and rat-anti-mouse CD19 (clone: 6D5, Biolegend), respectively, followed by using Alexa-488 conjugated goat-anti-rat IgG (clone: Poly4054, Biolegend). .. Immunohistochemistry was performed for the detection of MHC II molecules on lacrimal gland tissue cells by using rat-anti-mouse I-A/I-E antibody (clone: M5/114.15.2, Invitrogen) on cryosections. .. Gene expression profiling analysis Total RNA was isolated from murine lacrimal glands at week 0, 2, and 6 after immunization using TRIzol Plus RNA Purification Kit (Invitrogen).

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  • 97
    Thermo Fisher mouse anti rat mmp 9 monoclonal antibody
    Effect of irbesartan on changes in matrix metalloproteinase-9 expression induced by indomethacin in rat gastric mucosa. ( A ) Immunohistochemical staining of <t>MMP-9</t> in gastric sections (X 400). (a) Normal group showed no expression of MMP-9. (b) Irb group showed no expression of MMP-9. (c) Ind group showed strong expression of MMP-9 indicated by brown staining. (d) Ind + Ran group showed mild expression of MMP-9. (e) Ind + Irb group showed mild expression of MMP-9. ( B ) Quantification of MMP-9 staining as area percentage of immunopositive cells to the total area of microscopic field across 6 fields. Each bar with vertical line represents the mean ± S.E.M. of 6 fields. *vs normal, @ vs Ind (one-way ANOVA followed by Tukey’s multiple comparisons test; p
    Mouse Anti Rat Mmp 9 Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti rat mmp 9 monoclonal antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    Thermo Fisher anti rat mmp 9 monoclonal antibody
    Representative Western blot of <t>MMP-9</t> under non-reducing conditions. Lane 1: Day 0 plasma sample; Lane 2: Day 1 plasma sample; Lane 3: Day 2 plasma sample; Lane 4: Day 3 plasma sample; Lane 5: Day 4 plasma sample; Lane 6: Day 5 plasma sample; Lane 7: Day
    Anti Rat Mmp 9 Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rat mmp 9 monoclonal antibody/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    95
    Thermo Fisher rat anti α tubulin
    PLK1-independent furrow ingression requires centralspindlin. (A) Western blot of samples derived from HeLa cells transfected with either siLuc or siRACGAP1. The Western blot was probed with an anti-RACGAP1 antibody. <t>α-Tubulin</t> is shown as loading control. (B) HeLa cells were transfected with siLuc or siMKLP1 and processed for IF (right). The percentage of cells with detectable MKLP1 in anaphase was scored and used as a measure for knock-down efficiency (left). The number of cells that were scored per condition is indicated (n). One representative experiment out of two is shown. ****, P
    Rat Anti α Tubulin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti α tubulin/product/Thermo Fisher
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    86
    Thermo Fisher rat monoclonal anti mouse cd8
    Vaccination with KPC pulsed DCs resulted an antitumor T-cell response in KPC mice. (A) Flow cytometry for analyzing levels of surface proteins in immature dendritic cells (DC), mature DC or KPC loaded DC. A representative example of three independent experiments is shown. (B) Representative scatter plots of migrated DCs in the spleen and lymph node are shown. For DCs group, the spleen and pancreatic LN from the DCs treated mice were used. For control group, ndLN (inguinal) from the DCs treated mice and the spleen from untreated mice were used. (Representative experiment, n = 3). (C) In vivo CTLs assay. Naïve splenocytes were labeled with 20 × diluted CFSE (left peaks) or with CFSE (right peaks) and pulsed with UV-KPC cells, as target cells. Target cells at 5 × 10 6 were injected intravenously into the recipient mice 3-5 days after treatment. Splenocytes were collected from the recipient mice for detection of CFSE-labeled cells by flow cytometry at 16 hours. Quantitation of specific lysis are shown (n = 4). (D) Representative images of tumor infiltrating <t>CD8</t> + T cells. Scale bars represent 200 μm (inset, 25 μm) and (E) quantification are shown (n = 4). (F) Results of the proliferation of splenocytes. Spleen cells obtained from treated mice were co-cultured with UV-KPC, and the cell proliferation was evaluated by a CCK-8 kit assay. (G) Measurement of IFN-γ in serum from treated mice on day 4 after the last treatment (n = 5-6). (H) The production of IFN-γ in the culture supernatants. Splenocytes of surviving mice (DC treatment group and control group), were isolated when the mice were sacrificed and cultured in the presence of KPC tumor cells. Data were combined from three experiments. *P
    Rat Monoclonal Anti Mouse Cd8, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of irbesartan on changes in matrix metalloproteinase-9 expression induced by indomethacin in rat gastric mucosa. ( A ) Immunohistochemical staining of MMP-9 in gastric sections (X 400). (a) Normal group showed no expression of MMP-9. (b) Irb group showed no expression of MMP-9. (c) Ind group showed strong expression of MMP-9 indicated by brown staining. (d) Ind + Ran group showed mild expression of MMP-9. (e) Ind + Irb group showed mild expression of MMP-9. ( B ) Quantification of MMP-9 staining as area percentage of immunopositive cells to the total area of microscopic field across 6 fields. Each bar with vertical line represents the mean ± S.E.M. of 6 fields. *vs normal, @ vs Ind (one-way ANOVA followed by Tukey’s multiple comparisons test; p

    Journal: Scientific Reports

    Article Title: A Novel Role of Irbesartan in Gastroprotection against Indomethacin-Induced Gastric Injury in Rats: Targeting DDAH/ADMA and EGFR/ERK Signaling

    doi: 10.1038/s41598-018-22727-6

    Figure Lengend Snippet: Effect of irbesartan on changes in matrix metalloproteinase-9 expression induced by indomethacin in rat gastric mucosa. ( A ) Immunohistochemical staining of MMP-9 in gastric sections (X 400). (a) Normal group showed no expression of MMP-9. (b) Irb group showed no expression of MMP-9. (c) Ind group showed strong expression of MMP-9 indicated by brown staining. (d) Ind + Ran group showed mild expression of MMP-9. (e) Ind + Irb group showed mild expression of MMP-9. ( B ) Quantification of MMP-9 staining as area percentage of immunopositive cells to the total area of microscopic field across 6 fields. Each bar with vertical line represents the mean ± S.E.M. of 6 fields. *vs normal, @ vs Ind (one-way ANOVA followed by Tukey’s multiple comparisons test; p

    Article Snippet: The tissue sections were then incubated with mouse anti-rat MMP-9 monoclonal antibody (MA5-14228; Thermo Scientific, Rockford, IL, USA) for 60 minutes at 37 °C.

    Techniques: Expressing, Immunohistochemistry, Staining

    Representative Western blot of MMP-9 under non-reducing conditions. Lane 1: Day 0 plasma sample; Lane 2: Day 1 plasma sample; Lane 3: Day 2 plasma sample; Lane 4: Day 3 plasma sample; Lane 5: Day 4 plasma sample; Lane 6: Day 5 plasma sample; Lane 7: Day

    Journal: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society

    Article Title: Nitroglycerin Alters Matrix Remodeling Proteins in THP-1 Human Macrophages and Plasma Metalloproteinase Activity in Rats

    doi: 10.1016/j.niox.2010.12.002

    Figure Lengend Snippet: Representative Western blot of MMP-9 under non-reducing conditions. Lane 1: Day 0 plasma sample; Lane 2: Day 1 plasma sample; Lane 3: Day 2 plasma sample; Lane 4: Day 3 plasma sample; Lane 5: Day 4 plasma sample; Lane 6: Day 5 plasma sample; Lane 7: Day

    Article Snippet: The sources of other reagents were: RNA extraction kit, SV Total RNA® isolation system, from Promega (Madison, WI); RT2 PCR array system, CAPH-0166, RT2 PCR first strand kit from SABiosciences (Frederick, MD); Recombinant proMMP-9 protein and Quantikine human MMP-9, MMP-2, and TIMP-1 immunoassay kits from R & D system (Minneapolis, MN); 1,2- or 1,3-glyceryl dinitrate (GDN), and 1,2,4-butanetriol-1,4-dinitrate from Cerilliant (Round Rock, TX); Nuclear extraction kit from Chemicon International (Temecula, CA); NF-κB p50/p65 transcription factor assay from Millipore (Billerica, MA); Biotrak MMP-9 activity assay system from GE Healthcare (Piscataway, NJ); Alzet mini-osmotic pumps, model 2ML1, from DURECT Corporation (Cupertino, CA); One-Step Western™ kit for mouse, rabbit primary antibodies, and protein markers from GenScript Corporation (Piscataway, NJ); anti-rat MMP-9 monoclonal antibody from Thermo Fisher Scientific (Fermont, CA); CHEMICON® MMP Gelatinase Activity Assay Kit from Millipore (Billerica, MA); Sirocco protein precipitation plate from Waters (Milford, MA); 1,2- or 1,3-glyceryl dinitrate (GDN), and DualColor™ protein loading buffer from Fermentas (Glen Burnie, MD); recombinant active MMP-9 and MMP-9/ Lipocalin complex from Calbiochem (Gibbstown, NJ); Amersham full-range rainbow molecular weight markers from GE Healthcare (Piscataway, New Jersey), and anti-rat NGAL antibody from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Western Blot

    Comparison of MMP-9/NGAL, dimer protein, active MMP-9, and proMMP-9 protein expression changes in rat plasma samples of the rats implanted with osmotic pumps containing NTG stock solution. The band intensity obtained after Western blot analysis was normalized

    Journal: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society

    Article Title: Nitroglycerin Alters Matrix Remodeling Proteins in THP-1 Human Macrophages and Plasma Metalloproteinase Activity in Rats

    doi: 10.1016/j.niox.2010.12.002

    Figure Lengend Snippet: Comparison of MMP-9/NGAL, dimer protein, active MMP-9, and proMMP-9 protein expression changes in rat plasma samples of the rats implanted with osmotic pumps containing NTG stock solution. The band intensity obtained after Western blot analysis was normalized

    Article Snippet: The sources of other reagents were: RNA extraction kit, SV Total RNA® isolation system, from Promega (Madison, WI); RT2 PCR array system, CAPH-0166, RT2 PCR first strand kit from SABiosciences (Frederick, MD); Recombinant proMMP-9 protein and Quantikine human MMP-9, MMP-2, and TIMP-1 immunoassay kits from R & D system (Minneapolis, MN); 1,2- or 1,3-glyceryl dinitrate (GDN), and 1,2,4-butanetriol-1,4-dinitrate from Cerilliant (Round Rock, TX); Nuclear extraction kit from Chemicon International (Temecula, CA); NF-κB p50/p65 transcription factor assay from Millipore (Billerica, MA); Biotrak MMP-9 activity assay system from GE Healthcare (Piscataway, NJ); Alzet mini-osmotic pumps, model 2ML1, from DURECT Corporation (Cupertino, CA); One-Step Western™ kit for mouse, rabbit primary antibodies, and protein markers from GenScript Corporation (Piscataway, NJ); anti-rat MMP-9 monoclonal antibody from Thermo Fisher Scientific (Fermont, CA); CHEMICON® MMP Gelatinase Activity Assay Kit from Millipore (Billerica, MA); Sirocco protein precipitation plate from Waters (Milford, MA); 1,2- or 1,3-glyceryl dinitrate (GDN), and DualColor™ protein loading buffer from Fermentas (Glen Burnie, MD); recombinant active MMP-9 and MMP-9/ Lipocalin complex from Calbiochem (Gibbstown, NJ); Amersham full-range rainbow molecular weight markers from GE Healthcare (Piscataway, New Jersey), and anti-rat NGAL antibody from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Expressing, Western Blot

    3.3 MMP-9 protein expression and activity

    Journal: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society

    Article Title: Nitroglycerin Alters Matrix Remodeling Proteins in THP-1 Human Macrophages and Plasma Metalloproteinase Activity in Rats

    doi: 10.1016/j.niox.2010.12.002

    Figure Lengend Snippet: 3.3 MMP-9 protein expression and activity

    Article Snippet: The sources of other reagents were: RNA extraction kit, SV Total RNA® isolation system, from Promega (Madison, WI); RT2 PCR array system, CAPH-0166, RT2 PCR first strand kit from SABiosciences (Frederick, MD); Recombinant proMMP-9 protein and Quantikine human MMP-9, MMP-2, and TIMP-1 immunoassay kits from R & D system (Minneapolis, MN); 1,2- or 1,3-glyceryl dinitrate (GDN), and 1,2,4-butanetriol-1,4-dinitrate from Cerilliant (Round Rock, TX); Nuclear extraction kit from Chemicon International (Temecula, CA); NF-κB p50/p65 transcription factor assay from Millipore (Billerica, MA); Biotrak MMP-9 activity assay system from GE Healthcare (Piscataway, NJ); Alzet mini-osmotic pumps, model 2ML1, from DURECT Corporation (Cupertino, CA); One-Step Western™ kit for mouse, rabbit primary antibodies, and protein markers from GenScript Corporation (Piscataway, NJ); anti-rat MMP-9 monoclonal antibody from Thermo Fisher Scientific (Fermont, CA); CHEMICON® MMP Gelatinase Activity Assay Kit from Millipore (Billerica, MA); Sirocco protein precipitation plate from Waters (Milford, MA); 1,2- or 1,3-glyceryl dinitrate (GDN), and DualColor™ protein loading buffer from Fermentas (Glen Burnie, MD); recombinant active MMP-9 and MMP-9/ Lipocalin complex from Calbiochem (Gibbstown, NJ); Amersham full-range rainbow molecular weight markers from GE Healthcare (Piscataway, New Jersey), and anti-rat NGAL antibody from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Expressing, Activity Assay

    PLK1-independent furrow ingression requires centralspindlin. (A) Western blot of samples derived from HeLa cells transfected with either siLuc or siRACGAP1. The Western blot was probed with an anti-RACGAP1 antibody. α-Tubulin is shown as loading control. (B) HeLa cells were transfected with siLuc or siMKLP1 and processed for IF (right). The percentage of cells with detectable MKLP1 in anaphase was scored and used as a measure for knock-down efficiency (left). The number of cells that were scored per condition is indicated (n). One representative experiment out of two is shown. ****, P

    Journal: The Journal of Cell Biology

    Article Title: PLK1 plays dual roles in centralspindlin regulation during cytokinesis

    doi: 10.1083/jcb.201805036

    Figure Lengend Snippet: PLK1-independent furrow ingression requires centralspindlin. (A) Western blot of samples derived from HeLa cells transfected with either siLuc or siRACGAP1. The Western blot was probed with an anti-RACGAP1 antibody. α-Tubulin is shown as loading control. (B) HeLa cells were transfected with siLuc or siMKLP1 and processed for IF (right). The percentage of cells with detectable MKLP1 in anaphase was scored and used as a measure for knock-down efficiency (left). The number of cells that were scored per condition is indicated (n). One representative experiment out of two is shown. ****, P

    Article Snippet: Primary antibodies used were rabbit anti-KIF20A/MKLP2 (Bethyl [ITK]; A300-878A), rabbit anti-PRC1 (Santa Cruz; sc-8356), mouse anti–Aurora B (BD Transduction Laboratories; 611083), rabbit anti-Anillin , mouse anti-RhoA (Santa Cruz; sc-418), rabbit anti-MKLP1 (Santa Cruz; sc-867), rabbit anti-PLK1 (Santa Cruz; sc-17783), rat anti–α-tubulin (Thermo Fisher; MA1-80017), goat anti-RACGAP1 (Abcam; ab2270).

    Techniques: Western Blot, Derivative Assay, Transfection

    PRC1 depletion reveals PLK1-independent RhoA activation and furrow ingression. (A) Western blot of HeLa cells transfected with control siRNA (siLuc) or siRNA specific for PRC1. The Western blot was probed with an anti-PRC1 antibody. α-Tubulin is shown as loading control. (B) IF for PLK1, PRC1, and α-tubulin of HeLa cells transfected with siLuc or siPRC1. DNA was visualized using DAPI. (C) DIC stills of a live cell imaging experiment of HeLa cells transfected with either siLuc or siPRC1 with (PLK1 inh.) or without addition of BI2536 (100 nM) before anaphase onset. Time point 00:00 (hours:minutes) refers to the first frame where we observed separating sisters. Stills of more time frames are shown in Fig. S1 B. (D) Percentage of cells showing either complete furrow ingression, full-furrow ingression followed by furrow regression, visible but minimal furrow ingression, or no furrow ingression ( n = 100 cells imaged per condition). ****, P

    Journal: The Journal of Cell Biology

    Article Title: PLK1 plays dual roles in centralspindlin regulation during cytokinesis

    doi: 10.1083/jcb.201805036

    Figure Lengend Snippet: PRC1 depletion reveals PLK1-independent RhoA activation and furrow ingression. (A) Western blot of HeLa cells transfected with control siRNA (siLuc) or siRNA specific for PRC1. The Western blot was probed with an anti-PRC1 antibody. α-Tubulin is shown as loading control. (B) IF for PLK1, PRC1, and α-tubulin of HeLa cells transfected with siLuc or siPRC1. DNA was visualized using DAPI. (C) DIC stills of a live cell imaging experiment of HeLa cells transfected with either siLuc or siPRC1 with (PLK1 inh.) or without addition of BI2536 (100 nM) before anaphase onset. Time point 00:00 (hours:minutes) refers to the first frame where we observed separating sisters. Stills of more time frames are shown in Fig. S1 B. (D) Percentage of cells showing either complete furrow ingression, full-furrow ingression followed by furrow regression, visible but minimal furrow ingression, or no furrow ingression ( n = 100 cells imaged per condition). ****, P

    Article Snippet: Primary antibodies used were rabbit anti-KIF20A/MKLP2 (Bethyl [ITK]; A300-878A), rabbit anti-PRC1 (Santa Cruz; sc-8356), mouse anti–Aurora B (BD Transduction Laboratories; 611083), rabbit anti-Anillin , mouse anti-RhoA (Santa Cruz; sc-418), rabbit anti-MKLP1 (Santa Cruz; sc-867), rabbit anti-PLK1 (Santa Cruz; sc-17783), rat anti–α-tubulin (Thermo Fisher; MA1-80017), goat anti-RACGAP1 (Abcam; ab2270).

    Techniques: Activation Assay, Western Blot, Transfection, Live Cell Imaging

    PLK1 suppresses PRC1 to allow dynamic exchange of centralspindlin between the spindle midzone and cell cortex. (A) HeLa cells stably expressing the Tubby-Aurora B FRET sensor and H2B-mCherry were synchronized in G2 using RO3306 and imaged live after release from the Cdk1 inhibitor. 35 min after release, BI2536 (100 nM) was added. The emission ratio at the equatorial cortex was calculated for each time point (interval, 5 min). Mean ± SD of 11 cells is shown. (B) IF for α-tubulin and PRC1 of HeLa cells in anaphase transfected with the indicated siRNAs and treated with or without BI2536. (C) Representative stills of HeLa cells in anaphase with stable inducible expression of GFP::MKLP1 and RACGAP1::GFP and transfected with the indicated siRNAs and treated with or without BI2536 (100 nM). Numbers indicate the number of times the depicted localization was observed/total number of cells that was imaged live. See also Fig. S5 A. (D) Quantification of the IF intensity levels of PLK1, MKLP1, PRC1, and RACGAP1 at the spindle midzone in anaphase (see also Fig. S5 B). Each dot represents an individual cell. One representative experiment out of three is shown. Error bars depict SDs of the mean. ****, P

    Journal: The Journal of Cell Biology

    Article Title: PLK1 plays dual roles in centralspindlin regulation during cytokinesis

    doi: 10.1083/jcb.201805036

    Figure Lengend Snippet: PLK1 suppresses PRC1 to allow dynamic exchange of centralspindlin between the spindle midzone and cell cortex. (A) HeLa cells stably expressing the Tubby-Aurora B FRET sensor and H2B-mCherry were synchronized in G2 using RO3306 and imaged live after release from the Cdk1 inhibitor. 35 min after release, BI2536 (100 nM) was added. The emission ratio at the equatorial cortex was calculated for each time point (interval, 5 min). Mean ± SD of 11 cells is shown. (B) IF for α-tubulin and PRC1 of HeLa cells in anaphase transfected with the indicated siRNAs and treated with or without BI2536. (C) Representative stills of HeLa cells in anaphase with stable inducible expression of GFP::MKLP1 and RACGAP1::GFP and transfected with the indicated siRNAs and treated with or without BI2536 (100 nM). Numbers indicate the number of times the depicted localization was observed/total number of cells that was imaged live. See also Fig. S5 A. (D) Quantification of the IF intensity levels of PLK1, MKLP1, PRC1, and RACGAP1 at the spindle midzone in anaphase (see also Fig. S5 B). Each dot represents an individual cell. One representative experiment out of three is shown. Error bars depict SDs of the mean. ****, P

    Article Snippet: Primary antibodies used were rabbit anti-KIF20A/MKLP2 (Bethyl [ITK]; A300-878A), rabbit anti-PRC1 (Santa Cruz; sc-8356), mouse anti–Aurora B (BD Transduction Laboratories; 611083), rabbit anti-Anillin , mouse anti-RhoA (Santa Cruz; sc-418), rabbit anti-MKLP1 (Santa Cruz; sc-867), rabbit anti-PLK1 (Santa Cruz; sc-17783), rat anti–α-tubulin (Thermo Fisher; MA1-80017), goat anti-RACGAP1 (Abcam; ab2270).

    Techniques: Stable Transfection, Expressing, Transfection

    Vaccination with KPC pulsed DCs resulted an antitumor T-cell response in KPC mice. (A) Flow cytometry for analyzing levels of surface proteins in immature dendritic cells (DC), mature DC or KPC loaded DC. A representative example of three independent experiments is shown. (B) Representative scatter plots of migrated DCs in the spleen and lymph node are shown. For DCs group, the spleen and pancreatic LN from the DCs treated mice were used. For control group, ndLN (inguinal) from the DCs treated mice and the spleen from untreated mice were used. (Representative experiment, n = 3). (C) In vivo CTLs assay. Naïve splenocytes were labeled with 20 × diluted CFSE (left peaks) or with CFSE (right peaks) and pulsed with UV-KPC cells, as target cells. Target cells at 5 × 10 6 were injected intravenously into the recipient mice 3-5 days after treatment. Splenocytes were collected from the recipient mice for detection of CFSE-labeled cells by flow cytometry at 16 hours. Quantitation of specific lysis are shown (n = 4). (D) Representative images of tumor infiltrating CD8 + T cells. Scale bars represent 200 μm (inset, 25 μm) and (E) quantification are shown (n = 4). (F) Results of the proliferation of splenocytes. Spleen cells obtained from treated mice were co-cultured with UV-KPC, and the cell proliferation was evaluated by a CCK-8 kit assay. (G) Measurement of IFN-γ in serum from treated mice on day 4 after the last treatment (n = 5-6). (H) The production of IFN-γ in the culture supernatants. Splenocytes of surviving mice (DC treatment group and control group), were isolated when the mice were sacrificed and cultured in the presence of KPC tumor cells. Data were combined from three experiments. *P

    Journal: American Journal of Cancer Research

    Article Title: Dendritic cell immunotherapy induces anti-tumor effect in a transgenic mouse model of pancreatic ductal adenocarcinoma

    doi:

    Figure Lengend Snippet: Vaccination with KPC pulsed DCs resulted an antitumor T-cell response in KPC mice. (A) Flow cytometry for analyzing levels of surface proteins in immature dendritic cells (DC), mature DC or KPC loaded DC. A representative example of three independent experiments is shown. (B) Representative scatter plots of migrated DCs in the spleen and lymph node are shown. For DCs group, the spleen and pancreatic LN from the DCs treated mice were used. For control group, ndLN (inguinal) from the DCs treated mice and the spleen from untreated mice were used. (Representative experiment, n = 3). (C) In vivo CTLs assay. Naïve splenocytes were labeled with 20 × diluted CFSE (left peaks) or with CFSE (right peaks) and pulsed with UV-KPC cells, as target cells. Target cells at 5 × 10 6 were injected intravenously into the recipient mice 3-5 days after treatment. Splenocytes were collected from the recipient mice for detection of CFSE-labeled cells by flow cytometry at 16 hours. Quantitation of specific lysis are shown (n = 4). (D) Representative images of tumor infiltrating CD8 + T cells. Scale bars represent 200 μm (inset, 25 μm) and (E) quantification are shown (n = 4). (F) Results of the proliferation of splenocytes. Spleen cells obtained from treated mice were co-cultured with UV-KPC, and the cell proliferation was evaluated by a CCK-8 kit assay. (G) Measurement of IFN-γ in serum from treated mice on day 4 after the last treatment (n = 5-6). (H) The production of IFN-γ in the culture supernatants. Splenocytes of surviving mice (DC treatment group and control group), were isolated when the mice were sacrificed and cultured in the presence of KPC tumor cells. Data were combined from three experiments. *P

    Article Snippet: After blocking for 1 h at room temperature in blocking buffer (5% goat serum, 2.5% BSA in 1 × PBS), slides were incubated overnight in a humidified chamber at 4°C with anti-mouse CK19 (kindly provided by the Developmental Studies Hybridoma Bank), rabbit monoclonal anti-Ki67 (Clone SP6, Invitrogen), and rat monoclonal anti-mouse CD8 (Clone 4SM15, Invitrogen).

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, In Vivo, Labeling, Injection, Quantitation Assay, Lysis, Cell Culture, CCK-8 Assay, Isolation