peroxidase affinipure goat anti mouse igg fcγ subclass 2a specific  (Jackson Immuno)

 
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    Name:
    Peroxidase AffiniPure Goat Anti Mouse IgG Fcγ subclass 2a 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 IgG2a 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:
    115-035-206
    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 peroxidase affinipure goat anti mouse igg fcγ subclass 2a specific
    A27L-specific humoral immune response by ELISA. (A) Generation of total IgG antibody. (B) Generation of IgG1 and <t>IgG2a</t> antibodies. Sera from each group (n = 4) was collected two weeks after the last DNA immunization. Serum samples from each group of mice were diluted at 1:400 and incubated in a 96-well plate previously coated with recombinant VVWR A27. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments. A p value of less than 0.05 was considered significant, indicated by * = p
    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 IgG2a 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/peroxidase affinipure goat anti mouse igg fcγ subclass 2a 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 2a specific - by Bioz Stars, 2021-05
    93/100 stars

    Images

    1) Product Images from "Vaccination with a codon-optimized A27L-containing plasmid decreases virus replication and dissemination after vaccinia virus challenge"

    Article Title: Vaccination with a codon-optimized A27L-containing plasmid decreases virus replication and dissemination after vaccinia virus challenge

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2017.05.091

    A27L-specific humoral immune response by ELISA. (A) Generation of total IgG antibody. (B) Generation of IgG1 and IgG2a antibodies. Sera from each group (n = 4) was collected two weeks after the last DNA immunization. Serum samples from each group of mice were diluted at 1:400 and incubated in a 96-well plate previously coated with recombinant VVWR A27. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments. A p value of less than 0.05 was considered significant, indicated by * = p
    Figure Legend Snippet: A27L-specific humoral immune response by ELISA. (A) Generation of total IgG antibody. (B) Generation of IgG1 and IgG2a antibodies. Sera from each group (n = 4) was collected two weeks after the last DNA immunization. Serum samples from each group of mice were diluted at 1:400 and incubated in a 96-well plate previously coated with recombinant VVWR A27. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments. A p value of less than 0.05 was considered significant, indicated by * = p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay, Incubation, Recombinant

    2) Product Images from "Rabies virus-based COVID-19 vaccine CORAVAX™ induces high levels of neutralizing antibodies against SARS-CoV-2"

    Article Title: Rabies virus-based COVID-19 vaccine CORAVAX™ induces high levels of neutralizing antibodies against SARS-CoV-2

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-020-00248-6

    Analysis of the immune response to CORAVAX in mice. Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 IgG responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 IgG2/1 isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable IgG1 antibodies.
    Figure Legend Snippet: Analysis of the immune response to CORAVAX in mice. Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 IgG responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 IgG2/1 isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable IgG1 antibodies.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Recombinant Murine Growth Hormone Particles are More Immunogenic with Intravenous than Subcutaneous Administration"

    Article Title: Recombinant Murine Growth Hormone Particles are More Immunogenic with Intravenous than Subcutaneous Administration

    Journal: Journal of pharmaceutical sciences

    doi:

    Total IgG serum concentrations of pre-treatment (closed circles), SQ injected (open circles), IP injected (open squares) and IV injected (open triangles) mice throughout the study. Bars represent the average of each age group.
    Figure Legend Snippet: Total IgG serum concentrations of pre-treatment (closed circles), SQ injected (open circles), IP injected (open squares) and IV injected (open triangles) mice throughout the study. Bars represent the average of each age group.

    Techniques Used: Injection, Mouse Assay

    Anti-rmGH IgG isotype antibody titers for mice injected with ultra-centrifuged rmGH via the SQ route on day 22 (A), IP route on day 22 (B), IV route on day 22 (C), SQ route on day 36 (D), IP route on day 36 (E) and IV route on day 36 (F). No anti-rmGH
    Figure Legend Snippet: Anti-rmGH IgG isotype antibody titers for mice injected with ultra-centrifuged rmGH via the SQ route on day 22 (A), IP route on day 22 (B), IV route on day 22 (C), SQ route on day 36 (D), IP route on day 36 (E) and IV route on day 36 (F). No anti-rmGH

    Techniques Used: Mouse Assay, Injection

    Anti-rmGH IgG isotype antibody titers for mice injected with freeze-thawed rmGH via the SQ route on day 22 (A), IP route on day 22 (B), IV route on day 22 (C), SQ route on day 36 (D), IP route on day 36 (E) and IV route on day 36 (F). No anti-rmGH antibody
    Figure Legend Snippet: Anti-rmGH IgG isotype antibody titers for mice injected with freeze-thawed rmGH via the SQ route on day 22 (A), IP route on day 22 (B), IV route on day 22 (C), SQ route on day 36 (D), IP route on day 36 (E) and IV route on day 36 (F). No anti-rmGH antibody

    Techniques Used: Mouse Assay, Injection

    4) Product Images from "Soluble Spike DNA vaccine provides long-term protective immunity against SAR-CoV-2 in mice and nonhuman primates"

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

    Journal: bioRxiv

    doi: 10.1101/2020.10.09.334136

    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) .
    Figure Legend 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) .

    Techniques Used: 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.
    Figure Legend 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.

    Techniques Used: 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) .
    Figure Legend 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) .

    Techniques Used: 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.
    Figure Legend 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.

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

    5) Product Images from "The NS1 Glycoprotein Can Generate Dramatic Antibody-Enhanced Dengue Viral Replication in Normal Out-Bred Mice Resulting in Lethal Multi-Organ Disease"

    Article Title: The NS1 Glycoprotein Can Generate Dramatic Antibody-Enhanced Dengue Viral Replication in Normal Out-Bred Mice Resulting in Lethal Multi-Organ Disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0021024

    Immunoblot reactions of PAbs and MAbs against DENV-2 proteins. Approximately 1000 ng (odd numbered lanes) and 250 ng (even numbered lanes) concentrations of purified DENV-2 (NSx strain) virions, obtained from infected C6/36 supernatants, were heated at 100°C for 3 min and subjected to 9% (wt/vol) non-reduced SDS-PAGE and immuno-blotting. These strips were then reacted with either 1/200 dilutions of mouse PAbs generated against repeated infections with DENV-2 (NG-C strain) (lanes 1 and 2) or the pre-immunization (pre-i) and post-immunization (post-i) sera pooled from mice that had been generated against the purified NS1 glycoproteins of the DENV-2 16681 (pre-i: lanes 3 and 4; post-i: lanes 5 and 6), NG-C (pre-i: lanes 7 and 8; post-i: lanes 9 and 10) or NSx (pre-i: lanes 11 and 12; post-i: lanes 13 and 14) strains or ovalbumin (pre-i: lanes 15 and 16; post-i: lanes 17 and 18) or 1 µg/ml of IgG2a subclass MAbs specific for the DENV-2 NS1 (MAb 2A5.1) (lanes 19 and 20) the DENV E and prM glycoproteins (MAbs 2C5.1 and 2A4.1) (lanes 21 and 22) ( Table 1 ). The bound MAbs were then detected using a peroxidase-labelled goat anti-mouse IgG2a subclass-specific secondary PAbs and CND substrate. The locations of standard kilodalton (kD) molecular weight markers and the DENV-2 E (gp60 and 55), NS1 (gp48) and prM (gp20) glycoproteins are shown.
    Figure Legend Snippet: Immunoblot reactions of PAbs and MAbs against DENV-2 proteins. Approximately 1000 ng (odd numbered lanes) and 250 ng (even numbered lanes) concentrations of purified DENV-2 (NSx strain) virions, obtained from infected C6/36 supernatants, were heated at 100°C for 3 min and subjected to 9% (wt/vol) non-reduced SDS-PAGE and immuno-blotting. These strips were then reacted with either 1/200 dilutions of mouse PAbs generated against repeated infections with DENV-2 (NG-C strain) (lanes 1 and 2) or the pre-immunization (pre-i) and post-immunization (post-i) sera pooled from mice that had been generated against the purified NS1 glycoproteins of the DENV-2 16681 (pre-i: lanes 3 and 4; post-i: lanes 5 and 6), NG-C (pre-i: lanes 7 and 8; post-i: lanes 9 and 10) or NSx (pre-i: lanes 11 and 12; post-i: lanes 13 and 14) strains or ovalbumin (pre-i: lanes 15 and 16; post-i: lanes 17 and 18) or 1 µg/ml of IgG2a subclass MAbs specific for the DENV-2 NS1 (MAb 2A5.1) (lanes 19 and 20) the DENV E and prM glycoproteins (MAbs 2C5.1 and 2A4.1) (lanes 21 and 22) ( Table 1 ). The bound MAbs were then detected using a peroxidase-labelled goat anti-mouse IgG2a subclass-specific secondary PAbs and CND substrate. The locations of standard kilodalton (kD) molecular weight markers and the DENV-2 E (gp60 and 55), NS1 (gp48) and prM (gp20) glycoproteins are shown.

    Techniques Used: Purification, Infection, SDS Page, Generated, Mouse Assay, Molecular Weight

    Related Articles

    Amplification:

    Article Title: GUCY2C signaling opposes the acute radiation-induced GI syndrome
    Article Snippet: Fluorescent secondary antibodies were from Invitrogen. .. Tyramide signal amplification was used to detect GUCY2C and GUCA2A; secondary antibodies conjugated to horseradish peroxidase were from Jackson Immunoresearch Laboratories (cat #115-035-206 and #111-036-046, 1:1000 dilution), and fluorescein-conjugated tyramine was prepared from tyramine HCl (cat #T2879, Sigma) and NHS-fluorescein (cat #46410, Thermo Scientific) ( ). ..

    Incubation:

    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.

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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

    93
    Jackson Immuno peroxidase affinipure goat anti mouse igg fcγ subclass 2a specific
    Analysis of the immune response to CORAVAX in mice. Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 <t>IgG</t> responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 <t>IgG2/1</t> isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable <t>IgG1</t> antibodies.
    Peroxidase Affinipure Goat Anti Mouse Igg Fcγ Subclass 2a 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 2a 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 2a specific - by Bioz Stars, 2021-05
    93/100 stars
      Buy from Supplier

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    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

    Analysis of the immune response to CORAVAX in mice. Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 IgG responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 IgG2/1 isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable IgG1 antibodies.

    Journal: NPJ Vaccines

    Article Title: Rabies virus-based COVID-19 vaccine CORAVAX™ induces high levels of neutralizing antibodies against SARS-CoV-2

    doi: 10.1038/s41541-020-00248-6

    Figure Lengend Snippet: Analysis of the immune response to CORAVAX in mice. Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 IgG responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 IgG2/1 isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable IgG1 antibodies.

    Article Snippet: Plates were washed three times the next day, followed by the addition of goat anti-mouse-IgG-Fc HRP antibody (Southern Biotech, Cat# 1033-05, 1:8000 in PBST) or goat anti-mouse-IgG1-Fc HRP antibody (Jackson ImmunoResearch, Cat# 115-035-205, 1:8000 in PBST) or goat anti-mouse-IgG2a-Fc HRP antibody (Jackson ImmunoResearch, Cat# 115-035-206, 1:8000 in PBST) for 2 h at RT.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay