biotinylated rabbit anti goat igg  (Vector Laboratories)


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    Name:
    Biotinylated Rabbit Anti Goat IgG Antibody
    Description:
    Biotinylated Rabbit Anti Goat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Goat IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains The use of bovine products such as casein serum albumin or non fat dry milk as blocking agents in ELISA or blot assays may produce a high background due to cross reactivity of this antibody with bovine immunoglobulins that may be present If this antibody is to be used in these applications blocking with Animal Free Blocker solution Cat No SP 5030 is recommended This antibody is included in the VECTASTAIN ABC kits This antibody is suitable for use with goat sheep and bovine IgG primary antibodies
    Catalog Number:
    BA-5000
    Price:
    None
    Category:
    Antibodies
    Reactivity:
    Goat
    Size:
    1 5 mg
    Host:
    Rabbit
    Buy from Supplier


    Structured Review

    Vector Laboratories biotinylated rabbit anti goat igg
    Biotinylated Rabbit Anti Goat IgG Antibody
    Biotinylated Rabbit Anti Goat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Goat IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains The use of bovine products such as casein serum albumin or non fat dry milk as blocking agents in ELISA or blot assays may produce a high background due to cross reactivity of this antibody with bovine immunoglobulins that may be present If this antibody is to be used in these applications blocking with Animal Free Blocker solution Cat No SP 5030 is recommended This antibody is included in the VECTASTAIN ABC kits This antibody is suitable for use with goat sheep and bovine IgG primary antibodies
    https://www.bioz.com/result/biotinylated rabbit anti goat igg/product/Vector Laboratories
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated rabbit anti goat igg - by Bioz Stars, 2021-07
    98/100 stars

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    Related Articles

    Incubation:

    Article Title: Morphine Withdrawal Increases Glutamate Uptake and Surface Expression of Glutamate Transporter GLT1 at Hippocampal Synapses
    Article Snippet: .. Then the sections were incubated in antibodies against GLT1 (0.4 μg/ml) for 36 hr at 4°C, followed by biotinylated rabbit anti-goat IgG (1:200; Vector Laboratories, Burlingame, CA) and then stained with avidin–biotin–peroxidase complex (1:100; Vector ABC kit; Vector Laboratories). .. For the immunolabeling with gold–silver particles, rabbit antigoat IgG conjugated with 1.4 nm gold particles (1:100; Nanoprobes, Stony Brook, NY) were used to incubate for 2 hr and then enhanced with silver for 10 min (Nanoprobes).

    Article Title: Ghrelin Partially Protects Against Cisplatin-Induced Male Murine Gonadal Toxicity in a GHSR-1a-Dependent Manner 1
    Article Snippet: Tissue sections were deparaffinized, rehydrated, blocked with 10% normal rabbit serum, and incubated overnight with GATA-4 (1:200 of 200 μg/ml). .. After the slides were washed with 0.1% PBS-Tween, sections were incubated in biotinylated rabbit anti-goat antibody (1:200 dilution; Vector Laboratories, Burlingame, CA) for 1 h at room temperature. .. Stain was visualized by incubating with Vectastain ABC for 45 min followed by 3,3′-diaminobenzidine (Vector Laboratories) for 5 min and counterstaining with hematoxylin.

    Article Title: Differentiation of Retinal Ganglion Cells and Photoreceptor Precursors from Mouse Induced Pluripotent Stem Cells Carrying an Atoh7/Math5 Lineage Reporter
    Article Snippet: After washing, the cells were permeablized with 0.1% Triton X100 and incubated sequentially with primary and secondary antibodies to detect YFP and retinal cell type markers before FACS analyses using an LSRII flowcytometer (BD Biosciences). .. To detect YFP signals, after binding to primary rabbit anti-GFP antibodies, cells were incubated with biotinylated goat anti-rabbit antibodies (1∶200, Vector Laboratory) followed by Alexa 488-conjugated streptavidin (1∶500, Invitrogen). ..

    Article Title: Co-expression of Achaete-Scute Homologue-1 and Calcitonin Gene-Related Peptide during NNK-Induced Pulmonary Neuroendocrine Hyperplasia and Carcinogenesis in Hamsters
    Article Snippet: For brightfield double staining, sections were first incubated with mouse primary antibody against ASCL1, followed by biotinylated secondary antibody, ABC peroxidase and DAB (without intensification). .. Sections were then incubated with rabbit primary antibody against CGRP, followed by biotinylated secondary antibody, ABC alkaline phosphatase and Vector® Red (Vector Laboratories), with no subsequent counterstaining. .. For fluorescence studies, Alexa® Fluor 488 coupled goat anti-rabbit IgG and Alexa® Fluor 594 coupled goat anti-mouse IgG (Molecular Probes, Eugene, OR) were used as secondary antibodies (1:250) applied in solution together for 45 min at room temperature.

    Article Title: Systemic inflammation affects reperfusion following transient cerebral ischaemia
    Article Snippet: Sections were incubated overnight with primary antibodies as follows: mouse monoclonal anti-CD41/integrin alpha 2b antibody (platelets) (1:100, BD, NJ, USA), rabbit anti-Iba-1 (microglia) (1:1000, Abcam) and an immuno double-stain of rabbit anti-Iba-1 (microglia) (1:1000, Abcam) with goat anti-IL-1α (IL-1α co-localisation) (1:100 R & D Systems, USA), primary goat anti-mouse IL-1β IgG antibody (1:500, R & D Systems, Minneapolis, MN, USA). .. Sections were further incubated with secondary antibodies conjugated to Alexa 488 nm or Alexa 594 nm fluorochromes (1:500, Invitrogen, UK) for 2 h at room temperature or biotinylated rabbit anti-goat antibody (1:500, Vector Labs) for 2 h followed by nickel ammonia diaminobenzidine (DAB) staining procedure (for further details see ( , , , ) for IL-1 staining. .. A final wash was performed in PBS before application of Prolong Gold antifade reagent (with or without DAPI; Invitrogen, USA) used as the mounting medium for glass coverslips or DPX mounting medium (Agar Scientific, Stansted, UK).

    Staining:

    Article Title: Morphine Withdrawal Increases Glutamate Uptake and Surface Expression of Glutamate Transporter GLT1 at Hippocampal Synapses
    Article Snippet: .. Then the sections were incubated in antibodies against GLT1 (0.4 μg/ml) for 36 hr at 4°C, followed by biotinylated rabbit anti-goat IgG (1:200; Vector Laboratories, Burlingame, CA) and then stained with avidin–biotin–peroxidase complex (1:100; Vector ABC kit; Vector Laboratories). .. For the immunolabeling with gold–silver particles, rabbit antigoat IgG conjugated with 1.4 nm gold particles (1:100; Nanoprobes, Stony Brook, NY) were used to incubate for 2 hr and then enhanced with silver for 10 min (Nanoprobes).

    Article Title: Systemic inflammation affects reperfusion following transient cerebral ischaemia
    Article Snippet: Sections were incubated overnight with primary antibodies as follows: mouse monoclonal anti-CD41/integrin alpha 2b antibody (platelets) (1:100, BD, NJ, USA), rabbit anti-Iba-1 (microglia) (1:1000, Abcam) and an immuno double-stain of rabbit anti-Iba-1 (microglia) (1:1000, Abcam) with goat anti-IL-1α (IL-1α co-localisation) (1:100 R & D Systems, USA), primary goat anti-mouse IL-1β IgG antibody (1:500, R & D Systems, Minneapolis, MN, USA). .. Sections were further incubated with secondary antibodies conjugated to Alexa 488 nm or Alexa 594 nm fluorochromes (1:500, Invitrogen, UK) for 2 h at room temperature or biotinylated rabbit anti-goat antibody (1:500, Vector Labs) for 2 h followed by nickel ammonia diaminobenzidine (DAB) staining procedure (for further details see ( , , , ) for IL-1 staining. .. A final wash was performed in PBS before application of Prolong Gold antifade reagent (with or without DAPI; Invitrogen, USA) used as the mounting medium for glass coverslips or DPX mounting medium (Agar Scientific, Stansted, UK).

    Avidin-Biotin Assay:

    Article Title: Morphine Withdrawal Increases Glutamate Uptake and Surface Expression of Glutamate Transporter GLT1 at Hippocampal Synapses
    Article Snippet: .. Then the sections were incubated in antibodies against GLT1 (0.4 μg/ml) for 36 hr at 4°C, followed by biotinylated rabbit anti-goat IgG (1:200; Vector Laboratories, Burlingame, CA) and then stained with avidin–biotin–peroxidase complex (1:100; Vector ABC kit; Vector Laboratories). .. For the immunolabeling with gold–silver particles, rabbit antigoat IgG conjugated with 1.4 nm gold particles (1:100; Nanoprobes, Stony Brook, NY) were used to incubate for 2 hr and then enhanced with silver for 10 min (Nanoprobes).

    Article Title: Expression of orexin A and its receptor 1 in the human prostate
    Article Snippet: .. Goat polyclonal anti-OXA (sc-8070) and anti-OX1R antibodies (sc-8073), their respective blocking peptides (sc-8070 P, sc-8073 P), and mouse monoclonal anti-chromogranin A (Chr A) antibody (sc-47714) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); rabbit polyclonal anti-prepro-orexin antibody (AB3096), its blocking peptide (AG774) and monoclonal anti-tubulin antibody (MAB1637) from Chemicon International (Temecula, CA, USA); biotinylated rabbit anti-goat secondary antibodies and avidin–biotin complex (PK-6105) from Vector Laboratories (Burlingame, CA, USA); horseradish peroxidase-conjugated anti-goat IgG (A-5420) and anti-rabbit IgG (A-0545) from Sigma Chemical (St Louis, MO, USA). .. Triazol (15596-026) was purchased from Invitrogen (Carlsbad, CA, USA); the enhanced chemiluminescence (ECL) kit (RPN2209), the GFX PCR DNA and Gel Purification Kit (27-9602-01) from Amersham (Little Chalfont, UK); the DC protein assay kit (500-0111) from Bio-Rad Laboratories (Hercules, CA, USA); the primers for human prepro-orexin, OX1R and β-actin from Primm (Milan, Italy); the kit for PCR and RT-PCR (A1280) from Promega (Madison, WI, USA); and ethylenediamine tetra-acetic-acid (EDTA; 60-00-4) from Sigma Aldrich.

    Plasmid Preparation:

    Article Title: Morphine Withdrawal Increases Glutamate Uptake and Surface Expression of Glutamate Transporter GLT1 at Hippocampal Synapses
    Article Snippet: .. Then the sections were incubated in antibodies against GLT1 (0.4 μg/ml) for 36 hr at 4°C, followed by biotinylated rabbit anti-goat IgG (1:200; Vector Laboratories, Burlingame, CA) and then stained with avidin–biotin–peroxidase complex (1:100; Vector ABC kit; Vector Laboratories). .. For the immunolabeling with gold–silver particles, rabbit antigoat IgG conjugated with 1.4 nm gold particles (1:100; Nanoprobes, Stony Brook, NY) were used to incubate for 2 hr and then enhanced with silver for 10 min (Nanoprobes).

    Article Title: Co-expression of Achaete-Scute Homologue-1 and Calcitonin Gene-Related Peptide during NNK-Induced Pulmonary Neuroendocrine Hyperplasia and Carcinogenesis in Hamsters
    Article Snippet: For brightfield double staining, sections were first incubated with mouse primary antibody against ASCL1, followed by biotinylated secondary antibody, ABC peroxidase and DAB (without intensification). .. Sections were then incubated with rabbit primary antibody against CGRP, followed by biotinylated secondary antibody, ABC alkaline phosphatase and Vector® Red (Vector Laboratories), with no subsequent counterstaining. .. For fluorescence studies, Alexa® Fluor 488 coupled goat anti-rabbit IgG and Alexa® Fluor 594 coupled goat anti-mouse IgG (Molecular Probes, Eugene, OR) were used as secondary antibodies (1:250) applied in solution together for 45 min at room temperature.

    Binding Assay:

    Article Title: Differentiation of Retinal Ganglion Cells and Photoreceptor Precursors from Mouse Induced Pluripotent Stem Cells Carrying an Atoh7/Math5 Lineage Reporter
    Article Snippet: After washing, the cells were permeablized with 0.1% Triton X100 and incubated sequentially with primary and secondary antibodies to detect YFP and retinal cell type markers before FACS analyses using an LSRII flowcytometer (BD Biosciences). .. To detect YFP signals, after binding to primary rabbit anti-GFP antibodies, cells were incubated with biotinylated goat anti-rabbit antibodies (1∶200, Vector Laboratory) followed by Alexa 488-conjugated streptavidin (1∶500, Invitrogen). ..

    Blocking Assay:

    Article Title: Expression of orexin A and its receptor 1 in the human prostate
    Article Snippet: .. Goat polyclonal anti-OXA (sc-8070) and anti-OX1R antibodies (sc-8073), their respective blocking peptides (sc-8070 P, sc-8073 P), and mouse monoclonal anti-chromogranin A (Chr A) antibody (sc-47714) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); rabbit polyclonal anti-prepro-orexin antibody (AB3096), its blocking peptide (AG774) and monoclonal anti-tubulin antibody (MAB1637) from Chemicon International (Temecula, CA, USA); biotinylated rabbit anti-goat secondary antibodies and avidin–biotin complex (PK-6105) from Vector Laboratories (Burlingame, CA, USA); horseradish peroxidase-conjugated anti-goat IgG (A-5420) and anti-rabbit IgG (A-0545) from Sigma Chemical (St Louis, MO, USA). .. Triazol (15596-026) was purchased from Invitrogen (Carlsbad, CA, USA); the enhanced chemiluminescence (ECL) kit (RPN2209), the GFX PCR DNA and Gel Purification Kit (27-9602-01) from Amersham (Little Chalfont, UK); the DC protein assay kit (500-0111) from Bio-Rad Laboratories (Hercules, CA, USA); the primers for human prepro-orexin, OX1R and β-actin from Primm (Milan, Italy); the kit for PCR and RT-PCR (A1280) from Promega (Madison, WI, USA); and ethylenediamine tetra-acetic-acid (EDTA; 60-00-4) from Sigma Aldrich.

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  • 99
    Vector Laboratories biotinylated secondary goat anti mouse antibody
    Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects <t>biotinylated</t> Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).
    Biotinylated Secondary Goat Anti Mouse Antibody, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated secondary goat anti mouse antibody/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated secondary goat anti mouse antibody - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    99
    Vector Laboratories biotinylated goat anti rabbit igg secondary antibody
    ( A ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoblotting. ( B ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoprecipitation with beads conjugated to 4G10. Immunoprecipitates and whole-cell lysates (WCLs) were subjected to immunoblotting for phospho-tyrosines at the expected molecular mass of EGFR. ( C ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin, cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization, and the remaining surface biotin was cleaved. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture internalized <t>biotinylated</t> proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for tyrosine-phosphorylated EGFR. TS, total surface protein before stimulation. ( D ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin and cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization and degradation. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture all remaining biotinylated proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for total EGFR.
    Biotinylated Goat Anti Rabbit Igg Secondary Antibody, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated goat anti rabbit igg secondary antibody/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated goat anti rabbit igg secondary antibody - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    99
    Vector Laboratories biotinylated goat anti rabbit igg
    Detection of GNAQ in TG neurons during latency-reactivation cycle. TG were collected from 3 uninfected calves (U) (A), latently infected calves (L) (B), or latently infected calves treated with DEX for 6 h to initiate reactivation from latency (DEX) (C). Thin sections were cut from formalin-fixed, paraffin-embedded TG sections. The GNAQ antibody (ab75825; Abcam) was diluted 1:450. <t>Biotinylated</t> goat anti-rabbit <t>IgG</t> (Vector Laboratories) was used as a secondary antibody. Arrows denote GNAQ + neurons in the respective samples. (D) The percentage of GNAQ-positive neurons from 296 uninfected neurons, 241 latently infected neurons, and 209 TG neurons at 6 h after latently infected calves were treated with DEX. An asterisk denotes significant differences ( P
    Biotinylated Goat Anti Rabbit Igg, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated goat anti rabbit igg/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated goat anti rabbit igg - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

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    Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects biotinylated Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).

    Journal: PLoS ONE

    Article Title: Soluble amyloid-beta buffering by plaques in Alzheimer disease dementia versus high-pathology controls

    doi: 10.1371/journal.pone.0200251

    Figure Lengend Snippet: Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects biotinylated Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).

    Article Snippet: The following day, sections were washed 3x in TBS for 5 minutes each and incubated with a biotinylated secondary goat anti-mouse antibody at a 1:1000 dilution in TBS-X for 1 hour at room temperature (Vector Laboratories).

    Techniques: Enzyme-linked Immunosorbent Assay, Indirect ELISA, MANN-WHITNEY, Sandwich ELISA

    ( A ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoblotting. ( B ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoprecipitation with beads conjugated to 4G10. Immunoprecipitates and whole-cell lysates (WCLs) were subjected to immunoblotting for phospho-tyrosines at the expected molecular mass of EGFR. ( C ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin, cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization, and the remaining surface biotin was cleaved. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture internalized biotinylated proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for tyrosine-phosphorylated EGFR. TS, total surface protein before stimulation. ( D ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin and cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization and degradation. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture all remaining biotinylated proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for total EGFR.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: PI3K-p110α mediates the oncogenic activity induced by loss of the novel tumor suppressor PI3K-p85α

    doi: 10.1073/pnas.1704706114

    Figure Lengend Snippet: ( A ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoblotting. ( B ) Control and shPIK3R1 HMECs were starved and then stimulated with EGF (20 ng/mL). Protein lysates were collected and subjected to immunoprecipitation with beads conjugated to 4G10. Immunoprecipitates and whole-cell lysates (WCLs) were subjected to immunoblotting for phospho-tyrosines at the expected molecular mass of EGFR. ( C ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin, cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization, and the remaining surface biotin was cleaved. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture internalized biotinylated proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for tyrosine-phosphorylated EGFR. TS, total surface protein before stimulation. ( D ) Control and shPIK3R1 HMECs were starved; surface proteins were labeled with biotin and cells were stimulated with EGF (20 ng/mL) to initiate receptor internalization and degradation. Protein lysates were collected and subjected to streptavidin immunoprecipitation to capture all remaining biotinylated proteins. Immunoprecipitates and WCLs were subjected to immunoblotting for total EGFR.

    Article Snippet: Antibody signal was detected using biotinylated goat anti-rabbit IgG secondary antibody and DAB-developed (Vector Labs), and slides were counterstained with hematoxylin (Vector Labs).

    Techniques: Immunoprecipitation, Labeling

    Representative illustration of IL-1RII immunostaining in the human endometrium. Sections of endometrial tissue were incubated with mouse monoclonal anti-IL-1RII antibody ( A , proliferative day 13; B , secretory day 24; original magnification, ×68) or with an equivalent concentration of normal mouse IgGs ( C and D , respectively; original magnification, ×68). Sections were then incubated successively with biotinylated goat anti-mouse polyclonal antibody and avidin-biotinylated horseradish peroxidase complex. The immunoreaction was revealed with diaminobenzidine (brown staining) and hematoxylin was used for counterstaining (blue staining). Note the brown fine positive staining in stromal and epithelial cells (cellular staining) ( E–H ; original magnification, ×268), and the brown deposit ( arrow ) that is primarily located at the apical side of glandular ( E , secretory phase day 24) and surface ( F , secretory phase day 16) epithelium, or more spread within the glands lumen ( G , secretory phase day 16). Positive immunostaining is also detected in isolated stromal cells (c) ( G , secretory phase day 16) and microvessels (v) ( H , secretory phase day 24) found in the stroma in the secretory phase of the menstrual cycle. s = stroma, g = gland.

    Journal: The American Journal of Pathology

    Article Title: Decreased Expression of the Decoy Interleukin-1 Receptor Type II in Human Endometriosis

    doi:

    Figure Lengend Snippet: Representative illustration of IL-1RII immunostaining in the human endometrium. Sections of endometrial tissue were incubated with mouse monoclonal anti-IL-1RII antibody ( A , proliferative day 13; B , secretory day 24; original magnification, ×68) or with an equivalent concentration of normal mouse IgGs ( C and D , respectively; original magnification, ×68). Sections were then incubated successively with biotinylated goat anti-mouse polyclonal antibody and avidin-biotinylated horseradish peroxidase complex. The immunoreaction was revealed with diaminobenzidine (brown staining) and hematoxylin was used for counterstaining (blue staining). Note the brown fine positive staining in stromal and epithelial cells (cellular staining) ( E–H ; original magnification, ×268), and the brown deposit ( arrow ) that is primarily located at the apical side of glandular ( E , secretory phase day 24) and surface ( F , secretory phase day 16) epithelium, or more spread within the glands lumen ( G , secretory phase day 16). Positive immunostaining is also detected in isolated stromal cells (c) ( G , secretory phase day 16) and microvessels (v) ( H , secretory phase day 24) found in the stroma in the secretory phase of the menstrual cycle. s = stroma, g = gland.

    Article Snippet: After a PBS rinse, the sections were incubated for 60 minutes with a rabbit polyclonal anti-IL-1β antibody diluted 8:1,000 in PBS-1% bovine serum albumin (R and D Systems), washed in PBS, incubated for 60 minutes with a biotinylated goat anti-rabbit antibody (Vector Laboratories) diluted 1:100 in PBS-1% bovine serum albumin, washed again in PBS, and finally incubated simultaneously for 60 minutes in the dark with fluorescein isothiocyanate-conjugated streptavidin and a rhodamine-conjugated goat anti-mouse antibody (Sigma), which were used at a final dilution of 1:100 and 1:10 in PBS-1% bovine serum albumin, respectively.

    Techniques: Immunostaining, Incubation, Concentration Assay, Avidin-Biotin Assay, Staining, Isolation

    Detection of GNAQ in TG neurons during latency-reactivation cycle. TG were collected from 3 uninfected calves (U) (A), latently infected calves (L) (B), or latently infected calves treated with DEX for 6 h to initiate reactivation from latency (DEX) (C). Thin sections were cut from formalin-fixed, paraffin-embedded TG sections. The GNAQ antibody (ab75825; Abcam) was diluted 1:450. Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody. Arrows denote GNAQ + neurons in the respective samples. (D) The percentage of GNAQ-positive neurons from 296 uninfected neurons, 241 latently infected neurons, and 209 TG neurons at 6 h after latently infected calves were treated with DEX. An asterisk denotes significant differences ( P

    Journal: Journal of Virology

    Article Title: The Wnt Signaling Pathway Is Differentially Expressed during the Bovine Herpesvirus 1 Latency-Reactivation Cycle: Evidence That Two Protein Kinases Associated with Neuronal Survival, Akt3 and BMPR2, Are Expressed at Higher Levels during Latency

    doi: 10.1128/JVI.01937-17

    Figure Lengend Snippet: Detection of GNAQ in TG neurons during latency-reactivation cycle. TG were collected from 3 uninfected calves (U) (A), latently infected calves (L) (B), or latently infected calves treated with DEX for 6 h to initiate reactivation from latency (DEX) (C). Thin sections were cut from formalin-fixed, paraffin-embedded TG sections. The GNAQ antibody (ab75825; Abcam) was diluted 1:450. Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody. Arrows denote GNAQ + neurons in the respective samples. (D) The percentage of GNAQ-positive neurons from 296 uninfected neurons, 241 latently infected neurons, and 209 TG neurons at 6 h after latently infected calves were treated with DEX. An asterisk denotes significant differences ( P

    Article Snippet: The next day, slides were washed in 1× Tris-buffered saline (TBS) and incubated in biotinylated goat anti-rabbit IgG (PK-6101; Vector Laboratories) for 30 min at room temperature in a humidified chamber.

    Techniques: Infection, Formalin-fixed Paraffin-Embedded, Plasmid Preparation

    Akt3 is frequently detected in ORF2 + neurons during latency. Consecutive sections from formalin-fixed paraffin-embedded TG sections from latently infected calves were prepared. One section was stained with the Akt3 antibody (ab152157; Abcam) that was diluted 1:500. A consecutive section was stained with a peptide-specific ORF2 antibody (1:500 dilution). Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody for the sections. Areas of sections that contained ORF2 + neurons were subsequently examined for Akt3 staining. Numbers denote the ORF2-positive neurons, and neurons 1, 3, 4, and 5 were also Akt3 + . Magnification is approximately 400×, and these sections are representative of many sections that were examined.

    Journal: Journal of Virology

    Article Title: The Wnt Signaling Pathway Is Differentially Expressed during the Bovine Herpesvirus 1 Latency-Reactivation Cycle: Evidence That Two Protein Kinases Associated with Neuronal Survival, Akt3 and BMPR2, Are Expressed at Higher Levels during Latency

    doi: 10.1128/JVI.01937-17

    Figure Lengend Snippet: Akt3 is frequently detected in ORF2 + neurons during latency. Consecutive sections from formalin-fixed paraffin-embedded TG sections from latently infected calves were prepared. One section was stained with the Akt3 antibody (ab152157; Abcam) that was diluted 1:500. A consecutive section was stained with a peptide-specific ORF2 antibody (1:500 dilution). Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody for the sections. Areas of sections that contained ORF2 + neurons were subsequently examined for Akt3 staining. Numbers denote the ORF2-positive neurons, and neurons 1, 3, 4, and 5 were also Akt3 + . Magnification is approximately 400×, and these sections are representative of many sections that were examined.

    Article Snippet: The next day, slides were washed in 1× Tris-buffered saline (TBS) and incubated in biotinylated goat anti-rabbit IgG (PK-6101; Vector Laboratories) for 30 min at room temperature in a humidified chamber.

    Techniques: Formalin-fixed Paraffin-Embedded, Infection, Staining, Plasmid Preparation

    Comparison of Akt3 expression during the BoHV-1 latency-reactivation cycle. (A) TG were collected from 3 uninfected calves, 3 latently infected calves, or 3 latently infected calves treated with DEX for 6 h to initiate reactivation from latency. Thin sections were cut from formalin-fixed paraffin-embedded TG sections. The Akt3 antibody (ab152157; Abcam) was diluted 1:500. Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody. Blue arrows denote Akt3-positive TG neurons that contained an Akt3-positive nucleus and a counterstained nucleolus. Black arrows denote TG neurons in which the nucleus was not visible, but they were Akt3 + . Closed circles denote TG neurons that contain a nucleus in which the nucleolus is counterstained but was not stained by the Akt3 antibody. These images are representative of many sections stained with the Akt3 antibody. Magnification is approximately 400×. (B) The percentage of Akt3-positive TG neurons from 500 total neurons was estimated from sections derived from 3 latently infected calves, 3 mock-infected calves, and 3 latently infected calves treated with DEX for 6 h. An asterisk denotes significant differences ( P

    Journal: Journal of Virology

    Article Title: The Wnt Signaling Pathway Is Differentially Expressed during the Bovine Herpesvirus 1 Latency-Reactivation Cycle: Evidence That Two Protein Kinases Associated with Neuronal Survival, Akt3 and BMPR2, Are Expressed at Higher Levels during Latency

    doi: 10.1128/JVI.01937-17

    Figure Lengend Snippet: Comparison of Akt3 expression during the BoHV-1 latency-reactivation cycle. (A) TG were collected from 3 uninfected calves, 3 latently infected calves, or 3 latently infected calves treated with DEX for 6 h to initiate reactivation from latency. Thin sections were cut from formalin-fixed paraffin-embedded TG sections. The Akt3 antibody (ab152157; Abcam) was diluted 1:500. Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as a secondary antibody. Blue arrows denote Akt3-positive TG neurons that contained an Akt3-positive nucleus and a counterstained nucleolus. Black arrows denote TG neurons in which the nucleus was not visible, but they were Akt3 + . Closed circles denote TG neurons that contain a nucleus in which the nucleolus is counterstained but was not stained by the Akt3 antibody. These images are representative of many sections stained with the Akt3 antibody. Magnification is approximately 400×. (B) The percentage of Akt3-positive TG neurons from 500 total neurons was estimated from sections derived from 3 latently infected calves, 3 mock-infected calves, and 3 latently infected calves treated with DEX for 6 h. An asterisk denotes significant differences ( P

    Article Snippet: The next day, slides were washed in 1× Tris-buffered saline (TBS) and incubated in biotinylated goat anti-rabbit IgG (PK-6101; Vector Laboratories) for 30 min at room temperature in a humidified chamber.

    Techniques: Expressing, Infection, Formalin-fixed Paraffin-Embedded, Plasmid Preparation, Staining, Derivative Assay