secondary antibodies peroxidase conjugated affinipure goat anti mouse igg  (Jackson Immuno)

 
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    Name:
    Peroxidase AffiniPure Goat Anti Mouse IgG Fcγ subclass 1 specific
    Description:
    Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent The average molecular weight is reported to be about 160 kDa The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective Based on antigen binding assay and or ELISA the antibody reacts with the Fc portion of mouse IgG1 but not with other mouse IgG subclasses mouse IgM or the Fab portion of mouse immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with human bovine and rabbit serum proteins but it may cross react with immunoglobulins from other species
    Catalog Number:
    115-035-205
    Price:
    228.0
    Category:
    Anti Mouse IgG Subclass Specific Antibodies
    Conjugate:
    Horseradish Peroxidase
    Size:
    0 5 ml
    Format:
    Whole IgG
    Host:
    Goat
    Buy from Supplier


    Structured Review

    Jackson Immuno secondary antibodies peroxidase conjugated affinipure goat anti mouse igg
    Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent The average molecular weight is reported to be about 160 kDa The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective Based on antigen binding assay and or ELISA the antibody reacts with the Fc portion of mouse IgG1 but not with other mouse IgG subclasses mouse IgM or the Fab portion of mouse immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with human bovine and rabbit serum proteins but it may cross react with immunoglobulins from other species
    https://www.bioz.com/result/secondary antibodies peroxidase conjugated affinipure goat anti mouse igg/product/Jackson Immuno
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    secondary antibodies peroxidase conjugated affinipure goat anti mouse igg - by Bioz Stars, 2021-05
    93/100 stars

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    Incubation:

    Article Title: Immunolocalization of MMP9 and MMP2 in osteolytic metastasis originating from MDA-MB-231 human breast cancer cells
    Article Snippet: Subsequently, the sections were incubated with the following primary antibodies in BSA-PBS at room temperature for 2 h: SCGB2A2/mammaglobin A polyclonal antibody (MGB1; Proteintech; Sanying Biotechnology, Wuhan, China; cat. no. 235-1-AP; 1:50), anti-PCNA (Ab-1) mouse monoclonal antibody (PC10l; Epitomics; Abcam, Burlingame, CA, USA; cat. no. NA03; 1:50), mouse anti-human MMP2 monoclonal antibody (EMD Millipore, Billerica, MA, USA; cat. no. MAB3308; 1:50), mouse MMP9 antibody antigen affinifty-purified polyclonal goat IgG (EMD Millipore; cat. no. AF909; 1:50) and goat anti-MMP-13 polyclonal antibody (EMD Millipore; cat. no. AB8120; 1:50). .. Following rinsing with PBS, the sections were incubated with the following secondary antibodies for 1 h at room temperature: Polyclonal swine anti-rabbit immunoglobulin(Ig)/HRP from DakoCytomation, Denmark (cat. no. Nr.P 0399; 1:100), goat polyclonal anti-mouse IgG+IgM+IgA-H & L (HRP) from Abcam (cat. no. ab102448; 1:100), goat polyclonal anti-mouse IgG+IgM+IgA-H & L (HRP) from Abcam (cat. no. ab102448; 1:100), peroxidase-conjugated AffiniPure anti-goat++IgG (H+L) from Jackson Immunoresearch Laboratories, Inc. (West Grove, PA, USA; cat. no. 93894; 1:100) and peroxidase-conjugated AffiniPure anti-goat++IgG (H+L) from Jackson Immunoresearch Laboratories, Inc. (cat. no. 93894; 1:100). .. The immune complexes were then visualized using 3,3′-diamino-benzidine tetrahydrochloride (Sigma-Aldrich, St. Louis, MO, USA) as the substrate.

    Article Title: Human cytomegalovirus pp65 peptide-induced autoantibodies cross-reacts with TAF9 protein and induces lupus-like autoimmunity in BALB/c mice
    Article Snippet: For the competitive assays, 2 μg/well HCMVpp65422-439 , TAF9134-144 , dsDNA or TAF9 protein as competitor was mixed with 1 μg purified anti-HCMVpp65422-439 , anti-TAF9134-144 IgG, anti-TAF9 antibody or 100 μl eluted anti-TAF9 IgG fraction (1 ml/tube) with sterile PBS up to the final volume of 250 μl and added into peptide coating well for incubation at 37 °C for 2 hours. .. Following incubation, the microtiter plate was washed four times with TBST (TBS with 0.05% Tween-20) and any bound antibody was detected by horseradish peroxidase (HRP) conjugated anti-human/mouse IgG or anti-mouse IgG subclasses at 1:5,000 dilution (Jackson ImmunoResearch; Catalog code 109-035-088, 115-035-166, 115-035-205, 115-035-206, 115-035-207, 115-035-209) at 37 °C for 2 hours. .. After washing, TMB (Sigma-Aldrich) was used as the substrate, and HRP activity was measured at 450 nm by a microplate ELISA reader (EZ read 400).

    Article Title: Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates
    Article Snippet: Sera or BAL fluid were serially diluted in 5% SM, added to the wells and incubated for 2 hours at 37°C. .. Following incubation, plates were washed 5 times with 0.05% PBST and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG (Jackson ImmunoResearch Laboratories 115-035-003), IgG1 (Jackson ImmunoResearch Laboratories 115-035-205), or IgG2a (Jackson ImmunoResearch Laboratories 115-035-206) or IgG2b (Jackson ImmunoResearch Laboratories 115-035-207) for the mouse sera/BAL or anti-monkey IgG (Bethyl Laborabories A140-102P) for the NHP sera for 1 hour at 37°C. .. After final wash plates were developed using TMB solution (Surmodics TMBW-0100-01) and the reaction stopped with 2N H2 SO4.

    Article Title: Development of an anti‐BAG3 humanized antibody for treatment of pancreatic cancer
    Article Snippet: .. Plates were then extensively washed and incubated 30 minutes at room temperature with HRP‐conjugated anti‐mouse IgGs 1 : 2000 (115‐035‐205, Jackson ImmunoResearch, Cambridgeshire, UK) or anti‐human IgG 1 : 20 000 (A0170, Sigma‐Aldrich). .. Subsequently, TMB solution 1X (eBioscience, San Diego, CA, USA) was added to the wells for the analyte detection.

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    Jackson Immuno peroxidase affinipure goat anti mouse igg fcγ subclass 1 specific
    Antibody and T-cell responses after GX-19 vaccination in macaques. Macaques (n=3) were immunized with 3 mg of GX-19 as described in the methods. Serum and PBMCs were collected before (wk 0), during (wk 4, and 5.5) and after (wk 8) vaccination and were assessed for SARS-CoV-2 S-specific <t>IgG</t> antibodies by ELISA (A) and neutralizing antibodies against SARS-CoV-2 live-virus (B) . Data represent mean SEM of individual macaques, and dashed line indicate the assay limits of detection. The number of SARS-CoV-2 S-specific IFN-γ secreting cells in PBMCs was determined by IFN-γ ELISPOT assay after stimulation with peptide pools spanning the SARS-CoV-2 S protein. Shown are spot-forming cells (SFC) per 10 6 PBMCS in triplicate wells (C) . The frequency of S-specific CD4 + or CD8 + T cells producing IFN-γ, TNF-α, or IL-2 was determined by intracellular cytokine staining assays stimulated with SARS-CoV-2 S peptide pools. Shown are the frequency of S-specific CD4 + or CD8 + T cells after subtraction of background (DMSO vehicle) (D) .
    Peroxidase Affinipure Goat Anti Mouse Igg Fcγ Subclass 1 Specific, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase affinipure goat anti mouse igg fcγ subclass 1 specific/product/Jackson Immuno
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peroxidase affinipure goat anti mouse igg fcγ subclass 1 specific - by Bioz Stars, 2021-05
    93/100 stars
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    96
    Jackson Immuno peroxidase conjugated goat anti mouse igg
    Fig. 7. Binding of SNX17 and Dab1 to the intracellular domain of ApoER2. Microtiter plates (96-well) coated with a GST fusion protein containing the intracellular domain of ApoER2 were incubated ( A ) with the indicated amounts of purified <t>Dab1–MBP</t> expressed in E.coli or ( B ) with cell extracts derived from 293T cells expressing myc-tagged SNX17. Bound Dab1–MBP was visualized with the combination of an anti-MBP antibody and HRP-conjugated goat anti-rabbit <t>IgG.</t> Bound myc-SNX17 was visualized using the combination of an anti-myc antibody and HRP-conjugated goat anti-mouse IgG as described in Materials and methods. The color reaction was monitored by measuring absorption at 450 nm. ( C ) Competition of SNX17 binding by Dab1 was measured at two selected concentrations of myc-SNX17 (corresponding to 25 and 40% of cell lysate present in the incubation medium, respectively) in the absence and presence of 2 µg/ml Dab–MBP.
    Peroxidase Conjugated Goat Anti Mouse Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase conjugated goat anti mouse igg/product/Jackson Immuno
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peroxidase conjugated goat anti mouse igg - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

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    Antibody and T-cell responses after GX-19 vaccination in macaques. Macaques (n=3) were immunized with 3 mg of GX-19 as described in the methods. Serum and PBMCs were collected before (wk 0), during (wk 4, and 5.5) and after (wk 8) vaccination and were assessed for SARS-CoV-2 S-specific IgG antibodies by ELISA (A) and neutralizing antibodies against SARS-CoV-2 live-virus (B) . Data represent mean SEM of individual macaques, and dashed line indicate the assay limits of detection. The number of SARS-CoV-2 S-specific IFN-γ secreting cells in PBMCs was determined by IFN-γ ELISPOT assay after stimulation with peptide pools spanning the SARS-CoV-2 S protein. Shown are spot-forming cells (SFC) per 10 6 PBMCS in triplicate wells (C) . The frequency of S-specific CD4 + or CD8 + T cells producing IFN-γ, TNF-α, or IL-2 was determined by intracellular cytokine staining assays stimulated with SARS-CoV-2 S peptide pools. Shown are the frequency of S-specific CD4 + or CD8 + T cells after subtraction of background (DMSO vehicle) (D) .

    Journal: bioRxiv

    Article Title: Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates

    doi: 10.1101/2020.10.09.334136

    Figure Lengend Snippet: Antibody and T-cell responses after GX-19 vaccination in macaques. Macaques (n=3) were immunized with 3 mg of GX-19 as described in the methods. Serum and PBMCs were collected before (wk 0), during (wk 4, and 5.5) and after (wk 8) vaccination and were assessed for SARS-CoV-2 S-specific IgG antibodies by ELISA (A) and neutralizing antibodies against SARS-CoV-2 live-virus (B) . Data represent mean SEM of individual macaques, and dashed line indicate the assay limits of detection. The number of SARS-CoV-2 S-specific IFN-γ secreting cells in PBMCs was determined by IFN-γ ELISPOT assay after stimulation with peptide pools spanning the SARS-CoV-2 S protein. Shown are spot-forming cells (SFC) per 10 6 PBMCS in triplicate wells (C) . The frequency of S-specific CD4 + or CD8 + T cells producing IFN-γ, TNF-α, or IL-2 was determined by intracellular cytokine staining assays stimulated with SARS-CoV-2 S peptide pools. Shown are the frequency of S-specific CD4 + or CD8 + T cells after subtraction of background (DMSO vehicle) (D) .

    Article Snippet: Following incubation, plates were washed 5 times with 0.05% PBST and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG (Jackson ImmunoResearch Laboratories 115-035-003), IgG1 (Jackson ImmunoResearch Laboratories 115-035-205), or IgG2a (Jackson ImmunoResearch Laboratories 115-035-206) or IgG2b (Jackson ImmunoResearch Laboratories 115-035-207) for the mouse sera/BAL or anti-monkey IgG (Bethyl Laborabories A140-102P) for the NHP sera for 1 hour at 37°C.

    Techniques: Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Staining

    Immunization with GX-19 elicit Th1-biased T cell responses in mice. BALB/c mice (n=3-7/group) were immunized at week 0 and 2 with indicated doses of GX-19 or pGX27 (empty control vector) as described in the methods (A-C) . Sera were collected 2 weeks post-boost and assessed for SARS-CoV-2 S-specific IgG1 and IgG2a/b. Endpoint titers (A) , and endpoint tier ratios of IgG2a/b to IgG1 (B) were calculated. 2 weeks post-boost mouse splenocytes were isolated and re-stimulated with peptide pools spanning the SARS-CoV-2 S protein ex vivo . Indicated cytokines in the supernatants of culture were quantified using Th1/Th2 cytometric bead array kit (D) . T cell responses were measured by IFN-γ ELISPOT in splenocytes stimulated with peptide pools spanning the SARS-CoV-2 S protein (C) . Cells were stained for intracellular production of IFN-γ, TNF-α, and IL-2. Shown are the frequency of S-specific CD4 + or CD8 + T cells after subtraction of background (DMSO vehicle) (E) . Data representative of two independent experiments. P values determined by Mann-Whitney test.

    Journal: bioRxiv

    Article Title: Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates

    doi: 10.1101/2020.10.09.334136

    Figure Lengend Snippet: Immunization with GX-19 elicit Th1-biased T cell responses in mice. BALB/c mice (n=3-7/group) were immunized at week 0 and 2 with indicated doses of GX-19 or pGX27 (empty control vector) as described in the methods (A-C) . Sera were collected 2 weeks post-boost and assessed for SARS-CoV-2 S-specific IgG1 and IgG2a/b. Endpoint titers (A) , and endpoint tier ratios of IgG2a/b to IgG1 (B) were calculated. 2 weeks post-boost mouse splenocytes were isolated and re-stimulated with peptide pools spanning the SARS-CoV-2 S protein ex vivo . Indicated cytokines in the supernatants of culture were quantified using Th1/Th2 cytometric bead array kit (D) . T cell responses were measured by IFN-γ ELISPOT in splenocytes stimulated with peptide pools spanning the SARS-CoV-2 S protein (C) . Cells were stained for intracellular production of IFN-γ, TNF-α, and IL-2. Shown are the frequency of S-specific CD4 + or CD8 + T cells after subtraction of background (DMSO vehicle) (E) . Data representative of two independent experiments. P values determined by Mann-Whitney test.

    Article Snippet: Following incubation, plates were washed 5 times with 0.05% PBST and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG (Jackson ImmunoResearch Laboratories 115-035-003), IgG1 (Jackson ImmunoResearch Laboratories 115-035-205), or IgG2a (Jackson ImmunoResearch Laboratories 115-035-206) or IgG2b (Jackson ImmunoResearch Laboratories 115-035-207) for the mouse sera/BAL or anti-monkey IgG (Bethyl Laborabories A140-102P) for the NHP sera for 1 hour at 37°C.

    Techniques: Mouse Assay, Plasmid Preparation, Isolation, Ex Vivo, Enzyme-linked Immunospot, Staining, MANN-WHITNEY

    Diagram and immunogenicity of SARS-CoV-2 DNA vaccines. Schematic diagram of COVID-19 DNA vaccine expressing soluble SARS-CoV-2 S protein (S ΔTM ) or full-length SARS-CoV-2 S protein (S) (A) . BALB/c mice (n=4-10/group) were immunized at week 0 and 2 with pGX27-S ΔTM , pGX27-S or pGX27 (empty control vector) as described in the methods. Sera were collected 2 weeks post-prime (blue) and 2 weeks post-boost (red) and evaluated for SARS-CoV-2 S-specific IgG antibodies (B) .

    Journal: bioRxiv

    Article Title: Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates

    doi: 10.1101/2020.10.09.334136

    Figure Lengend Snippet: Diagram and immunogenicity of SARS-CoV-2 DNA vaccines. Schematic diagram of COVID-19 DNA vaccine expressing soluble SARS-CoV-2 S protein (S ΔTM ) or full-length SARS-CoV-2 S protein (S) (A) . BALB/c mice (n=4-10/group) were immunized at week 0 and 2 with pGX27-S ΔTM , pGX27-S or pGX27 (empty control vector) as described in the methods. Sera were collected 2 weeks post-prime (blue) and 2 weeks post-boost (red) and evaluated for SARS-CoV-2 S-specific IgG antibodies (B) .

    Article Snippet: Following incubation, plates were washed 5 times with 0.05% PBST and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG (Jackson ImmunoResearch Laboratories 115-035-003), IgG1 (Jackson ImmunoResearch Laboratories 115-035-205), or IgG2a (Jackson ImmunoResearch Laboratories 115-035-206) or IgG2b (Jackson ImmunoResearch Laboratories 115-035-207) for the mouse sera/BAL or anti-monkey IgG (Bethyl Laborabories A140-102P) for the NHP sera for 1 hour at 37°C.

    Techniques: Expressing, Mouse Assay, Plasmid Preparation

    GX-19 elicit robust binding and neutralizing antibody responses in mice. BALB/c mice (n=4-7/group) were immunized at week 0 and 2 with indicated doses of GX-19 or pGX27 as described in the methods (A-C) . Sera were collected 2 weeks post-prime (blue) and 2 weeks post-boost (red) and assessed for SARS-CoV-2 S-specific IgG antibodies by ELISA (A) , and for post-boost sera, neutralizing antibodies against SARS-CoV-2 live-virus (C) . BAL were collected 2 weeks post-boost and assayed for SARS-CoV-2 S-specific IgG antibodies by ELISA (B) . Data representative of two independent experiments. P values determined by Mann-Whitney test.

    Journal: bioRxiv

    Article Title: Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates

    doi: 10.1101/2020.10.09.334136

    Figure Lengend Snippet: GX-19 elicit robust binding and neutralizing antibody responses in mice. BALB/c mice (n=4-7/group) were immunized at week 0 and 2 with indicated doses of GX-19 or pGX27 as described in the methods (A-C) . Sera were collected 2 weeks post-prime (blue) and 2 weeks post-boost (red) and assessed for SARS-CoV-2 S-specific IgG antibodies by ELISA (A) , and for post-boost sera, neutralizing antibodies against SARS-CoV-2 live-virus (C) . BAL were collected 2 weeks post-boost and assayed for SARS-CoV-2 S-specific IgG antibodies by ELISA (B) . Data representative of two independent experiments. P values determined by Mann-Whitney test.

    Article Snippet: Following incubation, plates were washed 5 times with 0.05% PBST and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG (Jackson ImmunoResearch Laboratories 115-035-003), IgG1 (Jackson ImmunoResearch Laboratories 115-035-205), or IgG2a (Jackson ImmunoResearch Laboratories 115-035-206) or IgG2b (Jackson ImmunoResearch Laboratories 115-035-207) for the mouse sera/BAL or anti-monkey IgG (Bethyl Laborabories A140-102P) for the NHP sera for 1 hour at 37°C.

    Techniques: Binding Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Fig. 7. Binding of SNX17 and Dab1 to the intracellular domain of ApoER2. Microtiter plates (96-well) coated with a GST fusion protein containing the intracellular domain of ApoER2 were incubated ( A ) with the indicated amounts of purified Dab1–MBP expressed in E.coli or ( B ) with cell extracts derived from 293T cells expressing myc-tagged SNX17. Bound Dab1–MBP was visualized with the combination of an anti-MBP antibody and HRP-conjugated goat anti-rabbit IgG. Bound myc-SNX17 was visualized using the combination of an anti-myc antibody and HRP-conjugated goat anti-mouse IgG as described in Materials and methods. The color reaction was monitored by measuring absorption at 450 nm. ( C ) Competition of SNX17 binding by Dab1 was measured at two selected concentrations of myc-SNX17 (corresponding to 25 and 40% of cell lysate present in the incubation medium, respectively) in the absence and presence of 2 µg/ml Dab–MBP.

    Journal: The EMBO Journal

    Article Title: The PX-domain protein SNX17 interacts with members of the LDL receptor family and modulates endocytosis of the LDL receptor

    doi: 10.1093/emboj/cdf435

    Figure Lengend Snippet: Fig. 7. Binding of SNX17 and Dab1 to the intracellular domain of ApoER2. Microtiter plates (96-well) coated with a GST fusion protein containing the intracellular domain of ApoER2 were incubated ( A ) with the indicated amounts of purified Dab1–MBP expressed in E.coli or ( B ) with cell extracts derived from 293T cells expressing myc-tagged SNX17. Bound Dab1–MBP was visualized with the combination of an anti-MBP antibody and HRP-conjugated goat anti-rabbit IgG. Bound myc-SNX17 was visualized using the combination of an anti-myc antibody and HRP-conjugated goat anti-mouse IgG as described in Materials and methods. The color reaction was monitored by measuring absorption at 450 nm. ( C ) Competition of SNX17 binding by Dab1 was measured at two selected concentrations of myc-SNX17 (corresponding to 25 and 40% of cell lysate present in the incubation medium, respectively) in the absence and presence of 2 µg/ml Dab–MBP.

    Article Snippet: The following antibodies were obtained commercially from the sources indicated: anti-HA tag (16B12; Babco), anti-Myc (9E10, used as hybridoma supernatant from the corresponding cells; ATCC), anti-EEA1 (Affinity Bioreagents, Inc.), Oregon Green 488-labeled goat anti-mouse IgG (Molecular Probes), Alexa 594-labeled goat anti-rabbit IgG (Molecular Probes), goat anti-rabbit biotinylated IgG (Sigma), anti-MBP (E8030S; New England Biolabs), peroxidase- conjugated goat anti-mouse IgG (Jackson Immuno Research Laboratories) and peroxidase-conjugated anti-protein A (Sigma).

    Techniques: Binding Assay, Incubation, Purification, Derivative Assay, Expressing

    Target cell contact induces recruitment of 2B4 into DRM domains. Purified polyclonal human NK cells and 721.221 cells were either incubated separately (0′) or mixed (5′) for 5 min at 37°C. After cell lysis, DRM was isolated, neighboring fractions were combined and immunoprecipitated using a control antibody (IgG1) followed by anti-2B4 immunoprecipitation (2B4). Samples were analyzed by anti-2B4 Western blotting (2B4) and reblotted using an anti-phosphotyrosine antibody (2B4-P). DRM isolation was monitored by analyzing the different fraction using an anti-CD45 antibody and HRP-conjugated cholera toxin B-subunit (CTx).

    Journal: The Journal of Experimental Medicine

    Article Title: Natural Killer Cell Inhibitory Receptors Block Actin Cytoskeleton-dependent Recruitment of 2B4 (CD244) to Lipid Rafts

    doi: 10.1084/jem.20020427

    Figure Lengend Snippet: Target cell contact induces recruitment of 2B4 into DRM domains. Purified polyclonal human NK cells and 721.221 cells were either incubated separately (0′) or mixed (5′) for 5 min at 37°C. After cell lysis, DRM was isolated, neighboring fractions were combined and immunoprecipitated using a control antibody (IgG1) followed by anti-2B4 immunoprecipitation (2B4). Samples were analyzed by anti-2B4 Western blotting (2B4) and reblotted using an anti-phosphotyrosine antibody (2B4-P). DRM isolation was monitored by analyzing the different fraction using an anti-CD45 antibody and HRP-conjugated cholera toxin B-subunit (CTx).

    Article Snippet: Affinity-pure and HRP-conjugated goat anti–mouse IgG was purchased from Jackson ImmunoResearch Laboratories.

    Techniques: Purification, Incubation, Lysis, Isolation, Immunoprecipitation, Western Blot

    KIR2DL1 blocks DRM recruitment and phosphorylation of 2B4. YTS-2DL1 cells and 221-Cw3 or 221-Cw4 cells were either incubated separately (0′) or together (5′) for 5 min at 37°C. After cell lysis DRM was isolated, neighboring fractions were combined and analyzed by Western blotting using antibodies against 2B4 (top panel) or KIR2DL1 (bottom panel). DRM isolation was monitored by analyzing the different fraction using an anti-CD45 antibody and HRP-conjugated cholera toxin B-subunit as in Figs. 1 and 2 (unpublished data). (B) The DRM (4/5) and the soluble fractions (10/11) from the samples shown in panel A were immunoprecipitated (IP) using a control antibody (IgG1) followed by an anti-2B4 antibody (2B4) and analyzed by anti-2B4 Western blotting (2B4) and reblotted using an anti-phosphotyrosine antibody (2B4-P).

    Journal: The Journal of Experimental Medicine

    Article Title: Natural Killer Cell Inhibitory Receptors Block Actin Cytoskeleton-dependent Recruitment of 2B4 (CD244) to Lipid Rafts

    doi: 10.1084/jem.20020427

    Figure Lengend Snippet: KIR2DL1 blocks DRM recruitment and phosphorylation of 2B4. YTS-2DL1 cells and 221-Cw3 or 221-Cw4 cells were either incubated separately (0′) or together (5′) for 5 min at 37°C. After cell lysis DRM was isolated, neighboring fractions were combined and analyzed by Western blotting using antibodies against 2B4 (top panel) or KIR2DL1 (bottom panel). DRM isolation was monitored by analyzing the different fraction using an anti-CD45 antibody and HRP-conjugated cholera toxin B-subunit as in Figs. 1 and 2 (unpublished data). (B) The DRM (4/5) and the soluble fractions (10/11) from the samples shown in panel A were immunoprecipitated (IP) using a control antibody (IgG1) followed by an anti-2B4 antibody (2B4) and analyzed by anti-2B4 Western blotting (2B4) and reblotted using an anti-phosphotyrosine antibody (2B4-P).

    Article Snippet: Affinity-pure and HRP-conjugated goat anti–mouse IgG was purchased from Jackson ImmunoResearch Laboratories.

    Techniques: Incubation, Lysis, Isolation, Western Blot, Immunoprecipitation

    Characterization of mAbs against DENV1-4. (A) ELISA plates were coated with 1:800 dilutions of polyclonal rabbit anti-DENV hyper-immune sera at 4°C for 24 hours. After blocking, DENV1-4 viral supernatants were added, which were then detected by incubation with 1 μg/ml mAbs at RT for 2 hours. Next, the plates were incubated with HRP-conjugated goat anti-mouse IgG, and developed using OPD. The OD at a wavelength of 490 nm was measured. The cutoff values are represented by dotted lines. (B and C) BHK-21 cells were infected with DENV1-4, respectively. After 2 days, the infected cells were detected with mAbs. DD1-4, DD3-7, DD4-3, DD5-1, DD9-4, DD13-4, DD17-4, DD27-8, DD30-4, and DD31-3 are DENV4 serotype-specific mAbs. DD7-8, DD11-4, DD14-1, DD15-2, DD18-5, and DD33-2 are cross-reactive mAbs. 4G2 is the positive control.

    Journal: PLoS ONE

    Article Title: Generation of Monoclonal Antibodies against Dengue Virus Type 4 and Identification of Enhancing Epitopes on Envelope Protein

    doi: 10.1371/journal.pone.0136328

    Figure Lengend Snippet: Characterization of mAbs against DENV1-4. (A) ELISA plates were coated with 1:800 dilutions of polyclonal rabbit anti-DENV hyper-immune sera at 4°C for 24 hours. After blocking, DENV1-4 viral supernatants were added, which were then detected by incubation with 1 μg/ml mAbs at RT for 2 hours. Next, the plates were incubated with HRP-conjugated goat anti-mouse IgG, and developed using OPD. The OD at a wavelength of 490 nm was measured. The cutoff values are represented by dotted lines. (B and C) BHK-21 cells were infected with DENV1-4, respectively. After 2 days, the infected cells were detected with mAbs. DD1-4, DD3-7, DD4-3, DD5-1, DD9-4, DD13-4, DD17-4, DD27-8, DD30-4, and DD31-3 are DENV4 serotype-specific mAbs. DD7-8, DD11-4, DD14-1, DD15-2, DD18-5, and DD33-2 are cross-reactive mAbs. 4G2 is the positive control.

    Article Snippet: After washing, the membrane was incubated with HRP-conjugated goat anti-mouse IgG (Jackson ImmunoResearch Laboratories) and developed with chemiluminescence reagents (ECL; Amersham).

    Techniques: Enzyme-linked Immunosorbent Assay, Blocking Assay, Incubation, Infection, Positive Control