secondary antibodies  (Thermo Fisher)


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  • 95
    Name:
    F ab 2 Goat anti Mouse IgG Functional Grade Secondary Antibody
    Description:
    F ab 2 Goat anti Mouse IgG Functional Grade Secondary Antibody for Flow FN
    Catalog Number:
    16-5098-85
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
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    Structured Review

    Thermo Fisher secondary antibodies
    F ab 2 Goat anti Mouse IgG Functional Grade Secondary Antibody for Flow FN
    https://www.bioz.com/result/secondary antibodies/product/Thermo Fisher
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    secondary antibodies - by Bioz Stars, 2021-04
    95/100 stars

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    Related Articles

    Purification:

    Article Title: Heterophilic antibodies in sera from individuals without loxoscelism cross-react with phospholipase D from the venom of Loxosceles and Sicarius spiders
    Article Snippet: The purified IgG antibodies were evaluated by SDS-PAGE in gel at 10% and measuring absorbance at 280 nm in a TECAN® Infinite M200® PRO spectrofluorometer (Tecan Group Ltd., Männedorf, Switzerland). .. Subsequently, purified IgG antibodies were immunoselected using 2 μg of L. laeta venom adsorbed onto a nitrocellulose membrane in a 96-well Dot Blot Filtration Manifold system (Gibco BRL). .. The presence of the adsorbed proteins to the membrane was evaluated by staining with Ponceau red.

    Article Title: Heterophilic antibodies in sera from individuals without loxoscelism cross-react with phospholipase D from the venom of Loxosceles and Sicarius spiders
    Article Snippet: For this, 5 μL of serum from each individual was seeded into each well of the immunodiffusion plate and incubated at room temperature (22–25 °C) for 48 h. The measurement of each immunoprecipitation halo was performed using a ruler with accuracy of 0.01 mm, and the total IgG concentration was determined by comparison against data provided by the manufacturer (batch 1157, plate range: 201.8–3645.7 mg/dL; adult reference value: 710–1520 mg/dL). .. Purification of IgGs and immunoadsorption of antibodies against L. laeta venom Purification of IgG antibodies from sera was performed using the Pierce™ Protein G Agarose kit (Thermo Fisher Scientific, Inc. Waltham, MA, USA), following manufacturer’s instructions. .. Protein G Agarose resin in a 3:1 ratio with binding buffer (0.1 M sodium acetate, pH 5.0) was incubated with serum pools from groups 1 and 2, both previously diluted 1:1 in binding buffer and subsequently incubated in an orbital shaker at room temperature for 1 h and centrifuged at 500×g for 1 min. Then, each purification was washed twice with two volumes of PBS and again centrifuged at 500×g for 1 min, and the supernatant was discarded.

    Article Title: Entry of Francisella tularensis into Murine B Cells: The Role of B Cell Receptors and Complement Receptors
    Article Snippet: The cells were fixed with 3.8% formaldehyde for 30 min. After fixation, cells were washed three times in PBS, neutralized by incubation in PBS with 50 mM NH4 Cl for 10 min, permeabilized by 0.1% Triton X-100 for 5 min, and blocked by 3% bovine serum albumin (BSA) in PBS for 60 min at room temperature. .. Staining: cells were incubated for 45 min with the primary antibody against F . tularensis –purified immune high-titer anti-F . tularensis rabbit serum (subagglutinating ELISA titer 1:12800) diluted 1:3000 in 3% BSA in PBS and subsequently with secondary antibody goat anti-rabbit Alexa Fluor 488 (Molecular Probes, Grand Island, New York, USA) diluted 1:500 in 3% BSA in PBS. .. To detect LAMP-1, cells were stained with rat anti-mouse LAMP-1 antibody (Santa Cruz, Dallas, Texas, USA) diluted 1:100 in 3% BSA in PBS and then with secondary antibody donkey anti-rat Alexa Fluor 594 (Molecular Probes) diluted 1:500 in 3% BSA in PBS.

    Article Title: IgA Responses Following Recurrent Influenza Virus Vaccination
    Article Snippet: Purification of IgA and IgG AntibodiesTen subjects (five donors 18–35 y.o. and five donors 65–85 y.o.) with the highest HA-specific IgA titers post-vaccination were selected for isotype-specific antibody fractionation ( ). .. IgA1 and IgG antibodies were purified using jacalin agarose (Thermo Fisher Scientific, Waltham, MA, USA) and lectin beads (Thermo Fisher Scientific, Waltham, MA, USA) affinity chromatography, respectively. ..

    Dot Blot:

    Article Title: Heterophilic antibodies in sera from individuals without loxoscelism cross-react with phospholipase D from the venom of Loxosceles and Sicarius spiders
    Article Snippet: The purified IgG antibodies were evaluated by SDS-PAGE in gel at 10% and measuring absorbance at 280 nm in a TECAN® Infinite M200® PRO spectrofluorometer (Tecan Group Ltd., Männedorf, Switzerland). .. Subsequently, purified IgG antibodies were immunoselected using 2 μg of L. laeta venom adsorbed onto a nitrocellulose membrane in a 96-well Dot Blot Filtration Manifold system (Gibco BRL). .. The presence of the adsorbed proteins to the membrane was evaluated by staining with Ponceau red.

    Filtration:

    Article Title: Heterophilic antibodies in sera from individuals without loxoscelism cross-react with phospholipase D from the venom of Loxosceles and Sicarius spiders
    Article Snippet: The purified IgG antibodies were evaluated by SDS-PAGE in gel at 10% and measuring absorbance at 280 nm in a TECAN® Infinite M200® PRO spectrofluorometer (Tecan Group Ltd., Männedorf, Switzerland). .. Subsequently, purified IgG antibodies were immunoselected using 2 μg of L. laeta venom adsorbed onto a nitrocellulose membrane in a 96-well Dot Blot Filtration Manifold system (Gibco BRL). .. The presence of the adsorbed proteins to the membrane was evaluated by staining with Ponceau red.

    Co-Immunoprecipitation Assay:

    Article Title: The miR-361-3p increases enzalutamide (Enz) sensitivity via targeting the ARv7 and MKNK2 to better suppress the Enz-resistant prostate cancer
    Article Snippet: RNA immunoprecipitationCo-immunoprecipitation (Co-IP) of microRNA ribonucleoprotein complexes with anti-Ago2 or IgG was conducted. .. RNA Co-IP with anti-Ago2 or IgG antibodies was extracted using TRIzol reagent (Thermo Fisher Scientific) followed by detection of mRNA level through qRT-PCR. .. For detecting ARv7 3′UTR interaction with miRNA-361-3p, synthetic biotin-labeled sense or antisense ARv7 3′UTR was incubated with total RNA followed by detection of miRNA-361-3p levels in the pull-down assay.

    Quantitative RT-PCR:

    Article Title: The miR-361-3p increases enzalutamide (Enz) sensitivity via targeting the ARv7 and MKNK2 to better suppress the Enz-resistant prostate cancer
    Article Snippet: RNA immunoprecipitationCo-immunoprecipitation (Co-IP) of microRNA ribonucleoprotein complexes with anti-Ago2 or IgG was conducted. .. RNA Co-IP with anti-Ago2 or IgG antibodies was extracted using TRIzol reagent (Thermo Fisher Scientific) followed by detection of mRNA level through qRT-PCR. .. For detecting ARv7 3′UTR interaction with miRNA-361-3p, synthetic biotin-labeled sense or antisense ARv7 3′UTR was incubated with total RNA followed by detection of miRNA-361-3p levels in the pull-down assay.

    Recombinant:

    Article Title: Gain-of-function ADAMTS13 variants that are resistant to autoantibodies against ADAMTS13 in patients with acquired thrombotic thrombocytopenic purpura
    Article Snippet: A modified immunoprecipitation protocol plus Western blotting was used to detect the antigen-antibody interaction in solution as described previously., WT and variants (50 ng) were incubated 5 to 10 μL of normal human plasma or patient plasma and 30 μL of protein A/G-Sepharose 4B (Invitrogen) in 50mM Tris-HCl, pH 7.6, containing 0.15M NaCl, 1% BSA, 1% Triton X-100, and 0.1% Tween-20 at 4°C, overnight. .. After being washed with same buffer, bound recombinant WT and variants were eluted from the beads and determined by Western blotting with anti-V5 IgG (Invitrogen; 1:5000) in TBSTc, followed by IRDye 800CW-labeled goat anti–mouse IgG (1:20 000; LI-COR) as previously described. .. The interaction between ADAMTS13-spacer domain and VWF-A2 was modeled using the HHPred server plugin in PyMol software ( ).

    Article Title: Gain-of-function ADAMTS13 variants that are resistant to autoantibodies against ADAMTS13 in patients with acquired thrombotic thrombocytopenic purpura
    Article Snippet: The integrity of WT and variants in the concentrated conditioned medium were assessed by Western blotting after fractionation on 8% SDS-polyacrylamide gel under reduced conditions as described previously. .. After being transferred to a nitrocellulose membrane, recombinant WT and variants were blotted by anti-V5 IgG (Invitrogen; 1:5000) and IRdye800CW-labeled goat anti–mouse IgG (1:20 000; LI-COR) in 20mM Tris-HCl, 150mM NaCl containing 0.05% Tween-20 and 1% casein (TBSTc). .. The fluorescent signals obtained with an Odyssey imaging system (LI-COR) were converted to gray images.

    Western Blot:

    Article Title: Gain-of-function ADAMTS13 variants that are resistant to autoantibodies against ADAMTS13 in patients with acquired thrombotic thrombocytopenic purpura
    Article Snippet: A modified immunoprecipitation protocol plus Western blotting was used to detect the antigen-antibody interaction in solution as described previously., WT and variants (50 ng) were incubated 5 to 10 μL of normal human plasma or patient plasma and 30 μL of protein A/G-Sepharose 4B (Invitrogen) in 50mM Tris-HCl, pH 7.6, containing 0.15M NaCl, 1% BSA, 1% Triton X-100, and 0.1% Tween-20 at 4°C, overnight. .. After being washed with same buffer, bound recombinant WT and variants were eluted from the beads and determined by Western blotting with anti-V5 IgG (Invitrogen; 1:5000) in TBSTc, followed by IRDye 800CW-labeled goat anti–mouse IgG (1:20 000; LI-COR) as previously described. .. The interaction between ADAMTS13-spacer domain and VWF-A2 was modeled using the HHPred server plugin in PyMol software ( ).

    Staining:

    Article Title: Entry of Francisella tularensis into Murine B Cells: The Role of B Cell Receptors and Complement Receptors
    Article Snippet: The cells were fixed with 3.8% formaldehyde for 30 min. After fixation, cells were washed three times in PBS, neutralized by incubation in PBS with 50 mM NH4 Cl for 10 min, permeabilized by 0.1% Triton X-100 for 5 min, and blocked by 3% bovine serum albumin (BSA) in PBS for 60 min at room temperature. .. Staining: cells were incubated for 45 min with the primary antibody against F . tularensis –purified immune high-titer anti-F . tularensis rabbit serum (subagglutinating ELISA titer 1:12800) diluted 1:3000 in 3% BSA in PBS and subsequently with secondary antibody goat anti-rabbit Alexa Fluor 488 (Molecular Probes, Grand Island, New York, USA) diluted 1:500 in 3% BSA in PBS. .. To detect LAMP-1, cells were stained with rat anti-mouse LAMP-1 antibody (Santa Cruz, Dallas, Texas, USA) diluted 1:100 in 3% BSA in PBS and then with secondary antibody donkey anti-rat Alexa Fluor 594 (Molecular Probes) diluted 1:500 in 3% BSA in PBS.

    Incubation:

    Article Title: Entry of Francisella tularensis into Murine B Cells: The Role of B Cell Receptors and Complement Receptors
    Article Snippet: The cells were fixed with 3.8% formaldehyde for 30 min. After fixation, cells were washed three times in PBS, neutralized by incubation in PBS with 50 mM NH4 Cl for 10 min, permeabilized by 0.1% Triton X-100 for 5 min, and blocked by 3% bovine serum albumin (BSA) in PBS for 60 min at room temperature. .. Staining: cells were incubated for 45 min with the primary antibody against F . tularensis –purified immune high-titer anti-F . tularensis rabbit serum (subagglutinating ELISA titer 1:12800) diluted 1:3000 in 3% BSA in PBS and subsequently with secondary antibody goat anti-rabbit Alexa Fluor 488 (Molecular Probes, Grand Island, New York, USA) diluted 1:500 in 3% BSA in PBS. .. To detect LAMP-1, cells were stained with rat anti-mouse LAMP-1 antibody (Santa Cruz, Dallas, Texas, USA) diluted 1:100 in 3% BSA in PBS and then with secondary antibody donkey anti-rat Alexa Fluor 594 (Molecular Probes) diluted 1:500 in 3% BSA in PBS.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Entry of Francisella tularensis into Murine B Cells: The Role of B Cell Receptors and Complement Receptors
    Article Snippet: The cells were fixed with 3.8% formaldehyde for 30 min. After fixation, cells were washed three times in PBS, neutralized by incubation in PBS with 50 mM NH4 Cl for 10 min, permeabilized by 0.1% Triton X-100 for 5 min, and blocked by 3% bovine serum albumin (BSA) in PBS for 60 min at room temperature. .. Staining: cells were incubated for 45 min with the primary antibody against F . tularensis –purified immune high-titer anti-F . tularensis rabbit serum (subagglutinating ELISA titer 1:12800) diluted 1:3000 in 3% BSA in PBS and subsequently with secondary antibody goat anti-rabbit Alexa Fluor 488 (Molecular Probes, Grand Island, New York, USA) diluted 1:500 in 3% BSA in PBS. .. To detect LAMP-1, cells were stained with rat anti-mouse LAMP-1 antibody (Santa Cruz, Dallas, Texas, USA) diluted 1:100 in 3% BSA in PBS and then with secondary antibody donkey anti-rat Alexa Fluor 594 (Molecular Probes) diluted 1:500 in 3% BSA in PBS.

    Affinity Chromatography:

    Article Title: IgA Responses Following Recurrent Influenza Virus Vaccination
    Article Snippet: Purification of IgA and IgG AntibodiesTen subjects (five donors 18–35 y.o. and five donors 65–85 y.o.) with the highest HA-specific IgA titers post-vaccination were selected for isotype-specific antibody fractionation ( ). .. IgA1 and IgG antibodies were purified using jacalin agarose (Thermo Fisher Scientific, Waltham, MA, USA) and lectin beads (Thermo Fisher Scientific, Waltham, MA, USA) affinity chromatography, respectively. ..

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  • 94
    Thermo Fisher cadherin 11
    The distribution of <t>cadherin</t> 11 in the limbal area of the eye is illustrated. The section in A was stained with an antibody specific to cadherin 11 (green) and for DNA (blue). This staining is specific to the primary antibody for it is not observed in a section stained with the secondary antibody alone (B). The staining pattern of the trabecular meshwork is shown at higher magnification in C (arrow). A and B are taken at the same magnification and the scale bar in panel B represents 25 µm.
    Cadherin 11, 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
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    Average 94 stars, based on 1 article reviews
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    99
    Thermo Fisher cyclin d1
    eHSP90α had a stimulatory effect on HPDE cell anchorage independence but not cell proliferation. (A) Growth curves of HPDE cells treated with control medium (CTRL) or MϕCM for 1 to 5 days in the presence of control IgG or anti-CD91 antibody. Trypan blue exclusion assay was performed to count the numbers of viable cells. The data represent the averages of three independent experiments, and each experiment was performed in triplicate. (B) Growth curves of HPDE cells treated with PBS or 15 μg/ml of rHSP90α for 1 to 5 days. Trypan blue exclusion assay was performed, and the averages of three independent experiments are shown. (C) Flow cytometric analyses of the cell-cycle phase distribution of control medium (CTRL)- vs . MϕCM-treated HPDE cells (upper panel) and PBS- vs . rHSP90α-treated HPDE cells (lower panel). HPDE cells were treated for 24 h and trypsinized for ethanol fixation and propidium iodide staining. The histograph of cell-cycle phases was obtained from 10000 cells analyzed using a FACSCalibur flow cytometer. (D) The levels of <t>cyclin</t> D1, CDK4, Ser-780-phosphorylated Rb, Rb, cyclin E, cyclin A, Tyr-15-phosphorylated CDK2, CDK2, cyclin B, Tyr-15-phosphorylated CDC2, CDC2, and GAPDH in HPDE cells treated with PBS, 15 μg/ml rHSP90α, or 15 μg/ml rHSP90α plus preimmune IgG or anti-CD91 antibody. (E) Soft-agar colony-forming assay of PBS or rHSP90α-treated HPDE cells. HPDE cells (2000 cells seeded on the soft-agar layer in each well of a 6-well plate) were treated with or without 15 μg/ml rHSP90α and refreshed every 3 days for 3 weeks. The colonies were stained with Giemsa and photographed under a microscope (100 ×); only colonies consisting of ≥80 cells were counted. The mean ± SD values of three independent experiments are shown, and each experiment was performed in triplicate. # , P
    Cyclin D1, supplied by Thermo Fisher, 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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    Thermo Fisher igm
    B cell accumulation within the brain peaks by day 7 p.i Numbers of total CD19 + , <t>IgD</t> int <t>IgM</t> + , IgD − IgM + , and IgD − IgM − B cells determined by flow cytometry within the CLN (A) and brain (B) in naïve and infected mice at day 7, 14, and 21 p.i. Data are expressed as the mean number ± SEM and represent 3–4 independent experiments each comprising 3–6 pooled brains or CLNs per time point. B cells within brains of naïve mice were not detectable (ND). Significant differences between time points are denoted by * (p
    Igm, supplied by Thermo Fisher, 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


    The distribution of cadherin 11 in the limbal area of the eye is illustrated. The section in A was stained with an antibody specific to cadherin 11 (green) and for DNA (blue). This staining is specific to the primary antibody for it is not observed in a section stained with the secondary antibody alone (B). The staining pattern of the trabecular meshwork is shown at higher magnification in C (arrow). A and B are taken at the same magnification and the scale bar in panel B represents 25 µm.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Genomic Locus Modulating IOP in the BXD RI Mouse Strains

    doi: 10.1534/g3.118.200190

    Figure Lengend Snippet: The distribution of cadherin 11 in the limbal area of the eye is illustrated. The section in A was stained with an antibody specific to cadherin 11 (green) and for DNA (blue). This staining is specific to the primary antibody for it is not observed in a section stained with the secondary antibody alone (B). The staining pattern of the trabecular meshwork is shown at higher magnification in C (arrow). A and B are taken at the same magnification and the scale bar in panel B represents 25 µm.

    Article Snippet: The sections were incubated in primary antibodies (1:500) against Cadherin 11 (Thermofisher, Cat. #71-7600, Waltham, MA) overnight at 4°.

    Techniques: Staining

    eHSP90α had a stimulatory effect on HPDE cell anchorage independence but not cell proliferation. (A) Growth curves of HPDE cells treated with control medium (CTRL) or MϕCM for 1 to 5 days in the presence of control IgG or anti-CD91 antibody. Trypan blue exclusion assay was performed to count the numbers of viable cells. The data represent the averages of three independent experiments, and each experiment was performed in triplicate. (B) Growth curves of HPDE cells treated with PBS or 15 μg/ml of rHSP90α for 1 to 5 days. Trypan blue exclusion assay was performed, and the averages of three independent experiments are shown. (C) Flow cytometric analyses of the cell-cycle phase distribution of control medium (CTRL)- vs . MϕCM-treated HPDE cells (upper panel) and PBS- vs . rHSP90α-treated HPDE cells (lower panel). HPDE cells were treated for 24 h and trypsinized for ethanol fixation and propidium iodide staining. The histograph of cell-cycle phases was obtained from 10000 cells analyzed using a FACSCalibur flow cytometer. (D) The levels of cyclin D1, CDK4, Ser-780-phosphorylated Rb, Rb, cyclin E, cyclin A, Tyr-15-phosphorylated CDK2, CDK2, cyclin B, Tyr-15-phosphorylated CDC2, CDC2, and GAPDH in HPDE cells treated with PBS, 15 μg/ml rHSP90α, or 15 μg/ml rHSP90α plus preimmune IgG or anti-CD91 antibody. (E) Soft-agar colony-forming assay of PBS or rHSP90α-treated HPDE cells. HPDE cells (2000 cells seeded on the soft-agar layer in each well of a 6-well plate) were treated with or without 15 μg/ml rHSP90α and refreshed every 3 days for 3 weeks. The colonies were stained with Giemsa and photographed under a microscope (100 ×); only colonies consisting of ≥80 cells were counted. The mean ± SD values of three independent experiments are shown, and each experiment was performed in triplicate. # , P

    Journal: Oncoimmunology

    Article Title: Myeloid-derived macrophages and secreted HSP90α induce pancreatic ductal adenocarcinoma development

    doi: 10.1080/2162402X.2018.1424612

    Figure Lengend Snippet: eHSP90α had a stimulatory effect on HPDE cell anchorage independence but not cell proliferation. (A) Growth curves of HPDE cells treated with control medium (CTRL) or MϕCM for 1 to 5 days in the presence of control IgG or anti-CD91 antibody. Trypan blue exclusion assay was performed to count the numbers of viable cells. The data represent the averages of three independent experiments, and each experiment was performed in triplicate. (B) Growth curves of HPDE cells treated with PBS or 15 μg/ml of rHSP90α for 1 to 5 days. Trypan blue exclusion assay was performed, and the averages of three independent experiments are shown. (C) Flow cytometric analyses of the cell-cycle phase distribution of control medium (CTRL)- vs . MϕCM-treated HPDE cells (upper panel) and PBS- vs . rHSP90α-treated HPDE cells (lower panel). HPDE cells were treated for 24 h and trypsinized for ethanol fixation and propidium iodide staining. The histograph of cell-cycle phases was obtained from 10000 cells analyzed using a FACSCalibur flow cytometer. (D) The levels of cyclin D1, CDK4, Ser-780-phosphorylated Rb, Rb, cyclin E, cyclin A, Tyr-15-phosphorylated CDK2, CDK2, cyclin B, Tyr-15-phosphorylated CDC2, CDC2, and GAPDH in HPDE cells treated with PBS, 15 μg/ml rHSP90α, or 15 μg/ml rHSP90α plus preimmune IgG or anti-CD91 antibody. (E) Soft-agar colony-forming assay of PBS or rHSP90α-treated HPDE cells. HPDE cells (2000 cells seeded on the soft-agar layer in each well of a 6-well plate) were treated with or without 15 μg/ml rHSP90α and refreshed every 3 days for 3 weeks. The colonies were stained with Giemsa and photographed under a microscope (100 ×); only colonies consisting of ≥80 cells were counted. The mean ± SD values of three independent experiments are shown, and each experiment was performed in triplicate. # , P

    Article Snippet: The antibodies used were specific for human STAT3, phosphorylated STAT3 (EMD Millipore, Billerica, MA, USA), HSP90α (AbD Serotec), phosphorylated FAK (Invitrogen), phosphorylated CDK2 (Epitomics, Burlingame, CA, USA), phosphorylated Rb, Rb, phosphorylated CDC2 (Cell Signaling, Danvers, MA, USA), connexin-43 (Sigma, St. Louis, MO, USA), integrin αV (BD Biosciences, San Jose, CA, USA), cyclin D1, MMP-2, MMP-9 (Lab Vision/NeoMarkers Co., Fremont, CA, USA), CDK4, cyclin E, cyclin A, CDK2, cyclin B1, CDC2, FAK, TCF12, Twist-1, Snail, Slug, E-cadherin, connexin-26, fibronectin, and GAPDH (Santa Cruz Biotechnology).

    Techniques: Trypan Blue Exclusion Assay, Flow Cytometry, Staining, Cytometry, Microscopy

    B cell accumulation within the brain peaks by day 7 p.i Numbers of total CD19 + , IgD int IgM + , IgD − IgM + , and IgD − IgM − B cells determined by flow cytometry within the CLN (A) and brain (B) in naïve and infected mice at day 7, 14, and 21 p.i. Data are expressed as the mean number ± SEM and represent 3–4 independent experiments each comprising 3–6 pooled brains or CLNs per time point. B cells within brains of naïve mice were not detectable (ND). Significant differences between time points are denoted by * (p

    Journal: Brain, behavior, and immunity

    Article Title: Activated GL7+ B cells are maintained within the inflamed CNS in the absence of follicle formation during viral encephalomyelitis

    doi: 10.1016/j.bbi.2016.09.022

    Figure Lengend Snippet: B cell accumulation within the brain peaks by day 7 p.i Numbers of total CD19 + , IgD int IgM + , IgD − IgM + , and IgD − IgM − B cells determined by flow cytometry within the CLN (A) and brain (B) in naïve and infected mice at day 7, 14, and 21 p.i. Data are expressed as the mean number ± SEM and represent 3–4 independent experiments each comprising 3–6 pooled brains or CLNs per time point. B cells within brains of naïve mice were not detectable (ND). Significant differences between time points are denoted by * (p

    Article Snippet: Expression of cell surface markers was determined by staining with Ab specific for CD45 (30-F11; PerCP-Cy5.5), CD19 (1D3; PE-CF594), CD80 (16-10A1; PE), CD40 (3/23; PE), GL7 (FITC), CD95 (PE) (BD Biosciences, San Jose, CA), IgM (eB131-15F9; PE), IgD (11-26; APC), and CD38 (90; PE) (eBioscience, San Diego, CA).

    Techniques: Flow Cytometry, Cytometry, Infection, Mouse Assay

    IgM + and IgD + B cells are localized within the meninges and perivascular space Sagittal brain sections were stained with the indicated combinations of anti-B220, IgD, IgM, IgG, and laminin mAb to identify B cell localization to meninges and blood vessels and relative to basement membranes at day 7,14, and 21 p.i. (dpi) (A) B220 + (red) B cell localization relative to laminin + (green) structures. Arrows indicate isolated B cells emerging at parenchymal sites at day 14 p.i. (B) IgD + (red) B cells confined to the perivascular space. (C) IgM + (red) cells present within the laminin + subpial meningeal space. (D) Localization of both IgD + IgM + and IgM + IgD − B cells within the CNS at d14 p.i. (E) IgG + (red) B cells and laminin (green) at day 21 p.i. within the CNS. Representative images shown are z stack projected compilations from 2 independent experiments with 3 mice per experiment. (F) Percent distribution of isotype-unswitched (IgM + ) and isotype-switched (IgG+) B cells at day 14 and 21 p.i. For quantification, 6–8 Z stacks representing 2–3 animals per time point were counted manually for IgM+ or IgG+ B cells within the parenchyma or perivascular/meningeal space, defined by laminin staining. Data are expressed as the mean percentage ± SEM. Scale bars=50μm (A, E) and 20μm (B–D).

    Journal: Brain, behavior, and immunity

    Article Title: Activated GL7+ B cells are maintained within the inflamed CNS in the absence of follicle formation during viral encephalomyelitis

    doi: 10.1016/j.bbi.2016.09.022

    Figure Lengend Snippet: IgM + and IgD + B cells are localized within the meninges and perivascular space Sagittal brain sections were stained with the indicated combinations of anti-B220, IgD, IgM, IgG, and laminin mAb to identify B cell localization to meninges and blood vessels and relative to basement membranes at day 7,14, and 21 p.i. (dpi) (A) B220 + (red) B cell localization relative to laminin + (green) structures. Arrows indicate isolated B cells emerging at parenchymal sites at day 14 p.i. (B) IgD + (red) B cells confined to the perivascular space. (C) IgM + (red) cells present within the laminin + subpial meningeal space. (D) Localization of both IgD + IgM + and IgM + IgD − B cells within the CNS at d14 p.i. (E) IgG + (red) B cells and laminin (green) at day 21 p.i. within the CNS. Representative images shown are z stack projected compilations from 2 independent experiments with 3 mice per experiment. (F) Percent distribution of isotype-unswitched (IgM + ) and isotype-switched (IgG+) B cells at day 14 and 21 p.i. For quantification, 6–8 Z stacks representing 2–3 animals per time point were counted manually for IgM+ or IgG+ B cells within the parenchyma or perivascular/meningeal space, defined by laminin staining. Data are expressed as the mean percentage ± SEM. Scale bars=50μm (A, E) and 20μm (B–D).

    Article Snippet: Expression of cell surface markers was determined by staining with Ab specific for CD45 (30-F11; PerCP-Cy5.5), CD19 (1D3; PE-CF594), CD80 (16-10A1; PE), CD40 (3/23; PE), GL7 (FITC), CD95 (PE) (BD Biosciences, San Jose, CA), IgM (eB131-15F9; PE), IgD (11-26; APC), and CD38 (90; PE) (eBioscience, San Diego, CA).

    Techniques: Staining, Isolation, Mouse Assay

    Dynamics of B cell subsets within the CNS and CLN during JHMV infection Brain and CLN derived cells isolated from pooled organs of naïve or infected mice at day 7, 14, and 21 p.i. were analyzed for their IgD, IgM, and isotype-switched phenotype by flow cytometry. Representative density plots depict gating strategy for IgD + IgM + naïve B cells (1), IgD int IgM + early activated (2), IgD − IgM + activated (3), and IgD − IgM − isotype-switched (4) within CD45 hi CD19 + cells from brain and CLN as indicated. Red numbers indicate respective gates and black numbers show percent of populations. Stacked bar graphs show percentages of IgD int IgM + , IgD − IgM + , and IgD − IgM − cells within CD45 hi CD19 + cells and their changing dynamics over time. IgD + IgM + naïve B cells make up the remaining percentage of cells. B cells within brains of naïve mice were not detectable (ND). Data are expressed as the mean percentage ± SEM from 3–4 independent experiments each comprising 3–6 pooled brain or CLN per time point. Significant differences demarcated by * for IgD int IgM + , # for IgD − IgM + , and ¶ for IgD − IgM − B cell populations with * # ¶ denoting (p

    Journal: Brain, behavior, and immunity

    Article Title: Activated GL7+ B cells are maintained within the inflamed CNS in the absence of follicle formation during viral encephalomyelitis

    doi: 10.1016/j.bbi.2016.09.022

    Figure Lengend Snippet: Dynamics of B cell subsets within the CNS and CLN during JHMV infection Brain and CLN derived cells isolated from pooled organs of naïve or infected mice at day 7, 14, and 21 p.i. were analyzed for their IgD, IgM, and isotype-switched phenotype by flow cytometry. Representative density plots depict gating strategy for IgD + IgM + naïve B cells (1), IgD int IgM + early activated (2), IgD − IgM + activated (3), and IgD − IgM − isotype-switched (4) within CD45 hi CD19 + cells from brain and CLN as indicated. Red numbers indicate respective gates and black numbers show percent of populations. Stacked bar graphs show percentages of IgD int IgM + , IgD − IgM + , and IgD − IgM − cells within CD45 hi CD19 + cells and their changing dynamics over time. IgD + IgM + naïve B cells make up the remaining percentage of cells. B cells within brains of naïve mice were not detectable (ND). Data are expressed as the mean percentage ± SEM from 3–4 independent experiments each comprising 3–6 pooled brain or CLN per time point. Significant differences demarcated by * for IgD int IgM + , # for IgD − IgM + , and ¶ for IgD − IgM − B cell populations with * # ¶ denoting (p

    Article Snippet: Expression of cell surface markers was determined by staining with Ab specific for CD45 (30-F11; PerCP-Cy5.5), CD19 (1D3; PE-CF594), CD80 (16-10A1; PE), CD40 (3/23; PE), GL7 (FITC), CD95 (PE) (BD Biosciences, San Jose, CA), IgM (eB131-15F9; PE), IgD (11-26; APC), and CD38 (90; PE) (eBioscience, San Diego, CA).

    Techniques: Infection, Derivative Assay, Isolation, Mouse Assay, Flow Cytometry, Cytometry