human igg  (SouthernBiotech)

 
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  • 93
    Name:
    Goat Anti Human IgG AP
    Description:

    Catalog Number:
    2040-04
    Price:
    None
    Source:
    Pooled antisera from goats hyperimmunized with human IgG
    Applications:
    Quality tested applications for relevant formats include -ELISA 2-5FLISA 6Flow Cytometry 1,10,13,14Other referenced applications for relevant formats include -ELISpot 2,3,11Immunohistochemistry-Frozen Sections 7,8Immunohistochemistry-Paraffin Sections 9Immunocytochemistry 10-12Western Blot 4,12,17,18Immunoprecipitation 19Multiplex 3,15,16Depletion 14,20
    Format:
    AP (Alkaline Phosphatase)
    Isotype:
    Goat IgG
    Buy from Supplier


    Structured Review

    SouthernBiotech human igg
    Goat Anti Human IgG AP

    https://www.bioz.com/result/human igg/product/SouthernBiotech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human igg - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Eosinophils mediate tissue injury in autoimmune skin disease bullous pemphigoid"

    Article Title: Eosinophils mediate tissue injury in autoimmune skin disease bullous pemphigoid

    Journal: The Journal of investigative dermatology

    doi: 10.1016/j.jid.2017.11.031

    Anti-hNC16A IgE do not induce BP in neonatal hNC16A mice. Anti-hNC16A IgE recognized recombinant hNC16A by immunoblotting ( a, lane 2), stained the BMZ of hNC16A mouse skin sections by indirect IF ( b ), but failed to induce BP clinically and histologically in neonatal hNC16A mice at 48 hours post i.d. injection ( c ). ( d ) Immunostaining identified only neutrophils (PMN) in anti-hNC16A IgG-injected mouse skin; but no eosinophils were present in the skin of all anti-hNC16A antibody-injected mice. MPO and EPO enzymatic assays revealed significantly increased PMN in the anti-NC16A IgG-injected skin ( e ) but no eosinophil infiltration in the both anti-NC16A IgG- and IgE-injected skin 48 h post injection ( f ). Scale bars = 50 μm for panel b, scale bars = 100 μm for panels c, d. * p
    Figure Legend Snippet: Anti-hNC16A IgE do not induce BP in neonatal hNC16A mice. Anti-hNC16A IgE recognized recombinant hNC16A by immunoblotting ( a, lane 2), stained the BMZ of hNC16A mouse skin sections by indirect IF ( b ), but failed to induce BP clinically and histologically in neonatal hNC16A mice at 48 hours post i.d. injection ( c ). ( d ) Immunostaining identified only neutrophils (PMN) in anti-hNC16A IgG-injected mouse skin; but no eosinophils were present in the skin of all anti-hNC16A antibody-injected mice. MPO and EPO enzymatic assays revealed significantly increased PMN in the anti-NC16A IgG-injected skin ( e ) but no eosinophil infiltration in the both anti-NC16A IgG- and IgE-injected skin 48 h post injection ( f ). Scale bars = 50 μm for panel b, scale bars = 100 μm for panels c, d. * p

    Techniques Used: Mouse Assay, Recombinant, Staining, Injection, Immunostaining

    Anti-hNC16A IgE induce Eos infiltration but do not induce BP in adult hNC16A mice. Adult (8 week old) hNC16A mice were injected in the ear pinna and neonatal (24-36 h old) hNC16A mice were injected i.d. at dorsal back with anti-hNC16A IgE (100 ng/site) or anti-hNC16A IgG (100 μg/g body weight). Anti-hNC16A IgG induced infiltration of neutrophils ( a ) but not eosinophils ( c ) in both neonatal and adult mice and also triggered derma-epidermal separation in the IgG-injected ear ( e ). Anti-hNC16A IgE induced no neutrophil nor eosinophil infiltration in neonatal mice but increased neutrophil and eosinophil infiltration in adult mice at 24 and 48 time points ( b , d ). However, anti-hNC16A IgE-injected ear skin showed no derma-epidermal separation ( f ). Scale bars = 100 μm. 2-Way ANOVA, * p
    Figure Legend Snippet: Anti-hNC16A IgE induce Eos infiltration but do not induce BP in adult hNC16A mice. Adult (8 week old) hNC16A mice were injected in the ear pinna and neonatal (24-36 h old) hNC16A mice were injected i.d. at dorsal back with anti-hNC16A IgE (100 ng/site) or anti-hNC16A IgG (100 μg/g body weight). Anti-hNC16A IgG induced infiltration of neutrophils ( a ) but not eosinophils ( c ) in both neonatal and adult mice and also triggered derma-epidermal separation in the IgG-injected ear ( e ). Anti-hNC16A IgE induced no neutrophil nor eosinophil infiltration in neonatal mice but increased neutrophil and eosinophil infiltration in adult mice at 24 and 48 time points ( b , d ). However, anti-hNC16A IgE-injected ear skin showed no derma-epidermal separation ( f ). Scale bars = 100 μm. 2-Way ANOVA, * p

    Techniques Used: Mouse Assay, Injection

    2) Product Images from "Targeting of embryonic annexin A2 expressed on ovarian and breast cancer by the novel monoclonal antibody 2448"

    Article Title: Targeting of embryonic annexin A2 expressed on ovarian and breast cancer by the novel monoclonal antibody 2448

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24152

    Delivery of cytotoxic payloads by 2448 and ch2448 The potential of 2448 to be developed as an ADC was assessed. ( A ) Antibody 2448 was labelled with Cypher5e dye that maximally fluoresces in low pH environments (such as endosomal and lysosomal cellular compartments). At 4° C, IGROV1 cells were pre-incubated with dye-conjugated 2448 in the presence or absence of sodium azide (or with dye alone as a negative control). At T = 0 min, the temperature of samples was raised to 37° C. At T = 15 min, 2448 internalization was observed by an increase in fluorescence intensity. Less antibody internalization was observed on cells pre-treated with sodium azide as indicated by lower fluorescence intensity. ( B ) The cytotoxic effects of mAbs in-complex with secondary saporin conjugates (ZAP) was evaluated. Primary 2448 and ch2448 were pre-mixed with anti-mouse IgG (mAbZAP) and anti-human IgG saporin conjugates (HZAP), respectively. Ovarian and breast cancer cells were incubated with mixtures, primary antibody alone (2448 or ch2448), saporin conjugate alone (mAbZAP or HZAP) or buffer as a vehicle control. Post 72 h incubation, a significant cytotoxic effect was measured in IGROV1 and MCF7 cells that were treated with mAbs in-complex with secondary conjugates compared to cells treated with primary mAb or secondary conjugate alone. ( C ) IGROV1 and SKOV3 were treated with saporin-conjugated ch2448 (ch2448-saporin) or isotype control (IgG-saporin) at 30 mM. Cell viability was measured as a percentage of cells treated with buffer alone (% control). Post 72 h incubation, an 80% and 30% decrease in viability was observed on IGROV1 and SKOV3 cells, respectively. Cells treated with ch2448-saporin had shriveled unhealthy morphologies as observed by bright-field microscopy. Results were represented as the mean ± standard deviation of three independent experiments with triplicate. ( D ) Dose-dependent cytotoxicity of ch2448-saporin was evaluated. Post 72 h incubation, greater cytotoxicity was observed for ch2448-saporin than the IgG-saporin control. IC50 values of 6.59 nM and 17.43 nM were calculated on IGROV1 and SKOV3 cells, respectively. Cytotoxicity of free saporin was observed at higher concentrations ( > 0.1 µM). For (B) and (C), the relative cell viability was measured as a percentage of cells treated with buffer alone (% control). Results were represented as mean ± standard deviation of three independent experiments. Statistical significance was measured by a two-sided unpaired Student’s t -test ( * p
    Figure Legend Snippet: Delivery of cytotoxic payloads by 2448 and ch2448 The potential of 2448 to be developed as an ADC was assessed. ( A ) Antibody 2448 was labelled with Cypher5e dye that maximally fluoresces in low pH environments (such as endosomal and lysosomal cellular compartments). At 4° C, IGROV1 cells were pre-incubated with dye-conjugated 2448 in the presence or absence of sodium azide (or with dye alone as a negative control). At T = 0 min, the temperature of samples was raised to 37° C. At T = 15 min, 2448 internalization was observed by an increase in fluorescence intensity. Less antibody internalization was observed on cells pre-treated with sodium azide as indicated by lower fluorescence intensity. ( B ) The cytotoxic effects of mAbs in-complex with secondary saporin conjugates (ZAP) was evaluated. Primary 2448 and ch2448 were pre-mixed with anti-mouse IgG (mAbZAP) and anti-human IgG saporin conjugates (HZAP), respectively. Ovarian and breast cancer cells were incubated with mixtures, primary antibody alone (2448 or ch2448), saporin conjugate alone (mAbZAP or HZAP) or buffer as a vehicle control. Post 72 h incubation, a significant cytotoxic effect was measured in IGROV1 and MCF7 cells that were treated with mAbs in-complex with secondary conjugates compared to cells treated with primary mAb or secondary conjugate alone. ( C ) IGROV1 and SKOV3 were treated with saporin-conjugated ch2448 (ch2448-saporin) or isotype control (IgG-saporin) at 30 mM. Cell viability was measured as a percentage of cells treated with buffer alone (% control). Post 72 h incubation, an 80% and 30% decrease in viability was observed on IGROV1 and SKOV3 cells, respectively. Cells treated with ch2448-saporin had shriveled unhealthy morphologies as observed by bright-field microscopy. Results were represented as the mean ± standard deviation of three independent experiments with triplicate. ( D ) Dose-dependent cytotoxicity of ch2448-saporin was evaluated. Post 72 h incubation, greater cytotoxicity was observed for ch2448-saporin than the IgG-saporin control. IC50 values of 6.59 nM and 17.43 nM were calculated on IGROV1 and SKOV3 cells, respectively. Cytotoxicity of free saporin was observed at higher concentrations ( > 0.1 µM). For (B) and (C), the relative cell viability was measured as a percentage of cells treated with buffer alone (% control). Results were represented as mean ± standard deviation of three independent experiments. Statistical significance was measured by a two-sided unpaired Student’s t -test ( * p

    Techniques Used: Incubation, Negative Control, Fluorescence, Microscopy, Standard Deviation

    3) Product Images from "Spontaneous food allergy in Was−/− mice occurs independent of FcεRI-mediated mast cell activation"

    Article Title: Spontaneous food allergy in Was−/− mice occurs independent of FcεRI-mediated mast cell activation

    Journal: Allergy

    doi: 10.1111/all.13219

    Adjuvant-free oral sensitization in FcεRI-deficient animals. (A) Absence of surface on Was −/− Fcer1a −/− basophils. (B) Basophil numbers in Was −/− Fcer1a +/− (gray) and Was −/− Fcer1a −/− (black). (C) OVA-specific IgE and (D) IgG1 levels. (E) Serum MCPT1 in Was −/− Fcer1a +/− and Was −/− Fcer1a −/− littermates compared to OVA/alum-sensitized Fcer1a +/− or WT. Error depict as SEM. ***p
    Figure Legend Snippet: Adjuvant-free oral sensitization in FcεRI-deficient animals. (A) Absence of surface on Was −/− Fcer1a −/− basophils. (B) Basophil numbers in Was −/− Fcer1a +/− (gray) and Was −/− Fcer1a −/− (black). (C) OVA-specific IgE and (D) IgG1 levels. (E) Serum MCPT1 in Was −/− Fcer1a +/− and Was −/− Fcer1a −/− littermates compared to OVA/alum-sensitized Fcer1a +/− or WT. Error depict as SEM. ***p

    Techniques Used:

    Polysensitization in patients with food allergy and Wiskott-Aldrich syndrome. Antigen specificity profiling for common food allergens for (A) IgE and (B) IgG4 in serum from controls, food allergic (FA), and Wiskott Aldrich syndrome (WAS) patients.
    Figure Legend Snippet: Polysensitization in patients with food allergy and Wiskott-Aldrich syndrome. Antigen specificity profiling for common food allergens for (A) IgE and (B) IgG4 in serum from controls, food allergic (FA), and Wiskott Aldrich syndrome (WAS) patients.

    Techniques Used:

    4) Product Images from "Longitudinal Analysis of Levels of Immunoglobulins against BK Virus Capsid Proteins in Kidney Transplant Recipients ▿"

    Article Title: Longitudinal Analysis of Levels of Immunoglobulins against BK Virus Capsid Proteins in Kidney Transplant Recipients ▿

    Journal:

    doi: 10.1128/CVI.00206-08

    Correlation between immunoglobulin measurements and peak viral load in urine. Baseline OD (upper) showed a weak inverse correlation for anti-BK IgG, IgM, and IgA, but only the relationship with IgG OD was statistically significant ( r = −0.25;
    Figure Legend Snippet: Correlation between immunoglobulin measurements and peak viral load in urine. Baseline OD (upper) showed a weak inverse correlation for anti-BK IgG, IgM, and IgA, but only the relationship with IgG OD was statistically significant ( r = −0.25;

    Techniques Used:

    5) Product Images from "Biological Activities of Naturally Occurring Antibodies Reactive with Candida albicans Mannan "

    Article Title: Biological Activities of Naturally Occurring Antibodies Reactive with Candida albicans Mannan

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.1.209-218.2004

    Correlation between IgG, IgA, and IgM antimannan titers in sera from 34 normal adults. Antimannan titers were determined by ELISA (serotype A strain 36801 mannan) for each subject and a correlation was determined for the group of subjects.
    Figure Legend Snippet: Correlation between IgG, IgA, and IgM antimannan titers in sera from 34 normal adults. Antimannan titers were determined by ELISA (serotype A strain 36801 mannan) for each subject and a correlation was determined for the group of subjects.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Histogram showing distribution of mannan antibodies in sera of 34 normal adult donors. Levels of IgG, IgA, or IgM antibodies were determined by ELISA using serotype A strain 36801 mannan in the solid phase.
    Figure Legend Snippet: Histogram showing distribution of mannan antibodies in sera of 34 normal adult donors. Levels of IgG, IgA, or IgM antibodies were determined by ELISA using serotype A strain 36801 mannan in the solid phase.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    6) Product Images from "Human Immunoglobulin G2 (IgG2) and IgG4, but Not IgG1 or IgG3, Protect Mice against Cryptococcus neoformans Infection ▿"

    Article Title: Human Immunoglobulin G2 (IgG2) and IgG4, but Not IgG1 or IgG3, Protect Mice against Cryptococcus neoformans Infection ▿

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01161-06

    C1q binding by recombinant IgG against GXM detected by flow cytometry. C. neoformans was first incubated with the indicated isotypes, followed by fresh mouse serum. C1q was detected using rat anti-mouse C1q followed by rabbit F(ab′) 2 anti-rat IgG-FITC and analyzed by flow cytometry. Results for cryptococci incubated with anti-GXM are represented by a dark line with no fill, and results for those incubated with irrelevant anti-dansyl are represented by a light line and gray fill. Anti-GXM-coated C. neoformans incubated with heat-inactivated mouse serum gave results similar to those with control antibody (not shown).
    Figure Legend Snippet: C1q binding by recombinant IgG against GXM detected by flow cytometry. C. neoformans was first incubated with the indicated isotypes, followed by fresh mouse serum. C1q was detected using rat anti-mouse C1q followed by rabbit F(ab′) 2 anti-rat IgG-FITC and analyzed by flow cytometry. Results for cryptococci incubated with anti-GXM are represented by a dark line with no fill, and results for those incubated with irrelevant anti-dansyl are represented by a light line and gray fill. Anti-GXM-coated C. neoformans incubated with heat-inactivated mouse serum gave results similar to those with control antibody (not shown).

    Techniques Used: Binding Assay, Recombinant, Flow Cytometry, Cytometry, Incubation

    Binding patterns of recombinant human IgG subclasses to GXM. C. neoformans serotype D strain 24067 were incubated with recombinant IgGs followed by anti-human IgG(κ)-FITC. Stained organisms were then placed onto glass slides and examined by confocal microscopy at ×600 magnification. Shown at the top are control 2H1 and 13F1 mouse MAbs representative of annular (protective) and punctate (nonprotective) binding patterns, respectively, and detected with anti-mouse IgG(κ)-FITC.
    Figure Legend Snippet: Binding patterns of recombinant human IgG subclasses to GXM. C. neoformans serotype D strain 24067 were incubated with recombinant IgGs followed by anti-human IgG(κ)-FITC. Stained organisms were then placed onto glass slides and examined by confocal microscopy at ×600 magnification. Shown at the top are control 2H1 and 13F1 mouse MAbs representative of annular (protective) and punctate (nonprotective) binding patterns, respectively, and detected with anti-mouse IgG(κ)-FITC.

    Techniques Used: Binding Assay, Recombinant, Incubation, Staining, Confocal Microscopy

    7) Product Images from "Partial efficacy of a VSV-SIV/MVA-SIV vaccine regimen against oral SIV challenge in infant macaques"

    Article Title: Partial efficacy of a VSV-SIV/MVA-SIV vaccine regimen against oral SIV challenge in infant macaques

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2011.02.051

    Longitudinal SIV-specific plasma IgG and IgA antibodies. Panel A: Shown are the concentrations of IgA (Panel A) and IgA (Panel B) antibodies against SIV Env (left) and SIV Gag, Pol (right) proteins measured by ELISA in plasma of each vaccinated and unvaccinated
    Figure Legend Snippet: Longitudinal SIV-specific plasma IgG and IgA antibodies. Panel A: Shown are the concentrations of IgA (Panel A) and IgA (Panel B) antibodies against SIV Env (left) and SIV Gag, Pol (right) proteins measured by ELISA in plasma of each vaccinated and unvaccinated

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Correlation between SIV-specific antibodies and SIV replication levels. In Panel A and Panel B the correlation between peak plasma viremia and SIV Env-specific plasma IgA and IgG antibodies, respectively, at the time of challenge in vaccinated macaques
    Figure Legend Snippet: Correlation between SIV-specific antibodies and SIV replication levels. In Panel A and Panel B the correlation between peak plasma viremia and SIV Env-specific plasma IgA and IgG antibodies, respectively, at the time of challenge in vaccinated macaques

    Techniques Used:

    8) Product Images from "A Highly Expressing, Soluble, and Stable Plant-Made IgG Fusion Vaccine Strategy Enhances Antigen Immunogenicity in Mice Without Adjuvant"

    Article Title: A Highly Expressing, Soluble, and Stable Plant-Made IgG Fusion Vaccine Strategy Enhances Antigen Immunogenicity in Mice Without Adjuvant

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.576012

    Modified IgG fusions have improved expression and solubility (A) . ELISA and gel quantification of IgG fusion construct expression. Clarified protein extracts (“soluble” fraction) from leaf spots agroinfiltrated with each IgG fusion construct were analyzed by either ELISA, or SDS-PAGE followed by gel image quantification. For ZE3 ELISA, plates were coated with polyclonal mouse anti-ZE3, incubated with serial dilutions of extracts from each IgG fusion using purified HLZ as a standard, and probed with goat anti-human IgG-HRP. For IgG ELISA, plates were coated with serial dilutions of extracts or human IgG standard and probed with goat anti-human IgG-HRP. For gel quantification, ImageJ software was used to compare the IgG fusion band intensity visualized on stain-free polyacrylamide gels using purified 6D8 antibody as standard. Columns represent means ± standard error from three independently infiltrated leaf samples (B) . Clarified leaf extracts were separated by reducing SDS-PAGE and a representative gel image is shown. The general band position corresponding to each respective heavy chain/ZE3 fusion is indicated “ZH/HZ.” The small shift in size in HLZe (~67 kDa of the reduced heavy chain fusion before glycosylation) and HLZd (~65 kDa) or HLZ (~64.5 kDa) is due to epitope tag truncation. ZH and ZHx have a predicted size of ~64.5 kDa. The “
    Figure Legend Snippet: Modified IgG fusions have improved expression and solubility (A) . ELISA and gel quantification of IgG fusion construct expression. Clarified protein extracts (“soluble” fraction) from leaf spots agroinfiltrated with each IgG fusion construct were analyzed by either ELISA, or SDS-PAGE followed by gel image quantification. For ZE3 ELISA, plates were coated with polyclonal mouse anti-ZE3, incubated with serial dilutions of extracts from each IgG fusion using purified HLZ as a standard, and probed with goat anti-human IgG-HRP. For IgG ELISA, plates were coated with serial dilutions of extracts or human IgG standard and probed with goat anti-human IgG-HRP. For gel quantification, ImageJ software was used to compare the IgG fusion band intensity visualized on stain-free polyacrylamide gels using purified 6D8 antibody as standard. Columns represent means ± standard error from three independently infiltrated leaf samples (B) . Clarified leaf extracts were separated by reducing SDS-PAGE and a representative gel image is shown. The general band position corresponding to each respective heavy chain/ZE3 fusion is indicated “ZH/HZ.” The small shift in size in HLZe (~67 kDa of the reduced heavy chain fusion before glycosylation) and HLZd (~65 kDa) or HLZ (~64.5 kDa) is due to epitope tag truncation. ZH and ZHx have a predicted size of ~64.5 kDa. The “

    Techniques Used: Modification, Expressing, Solubility, Enzyme-linked Immunosorbent Assay, Construct, SDS Page, Incubation, Purification, Software, Staining

    Stability and C1q binding of purified IgG fusions. Samples of purified IgG fusions were frozen and thawed once after purification (initial), or additionally subjected to either additional five freeze/thaw cycles, incubation for 2 weeks at 4°C, or incubation for 2 weeks at 23˚C (A) . After each treatment, samples were separated on reducing and nonreducing SDS-PAGE gels, and the relative proportion of fully assembled product was analyzed using ImageJ software. The initial measurement reflects degradation which occurred during expression or purification, whereas the other measurements reflect degradation which occurred during each respective treatment (B) . Samples from each treatment were analyzed by C1q binding ELISA. Columns represent the mean OD 450 value ± standard error from three samples.
    Figure Legend Snippet: Stability and C1q binding of purified IgG fusions. Samples of purified IgG fusions were frozen and thawed once after purification (initial), or additionally subjected to either additional five freeze/thaw cycles, incubation for 2 weeks at 4°C, or incubation for 2 weeks at 23˚C (A) . After each treatment, samples were separated on reducing and nonreducing SDS-PAGE gels, and the relative proportion of fully assembled product was analyzed using ImageJ software. The initial measurement reflects degradation which occurred during expression or purification, whereas the other measurements reflect degradation which occurred during each respective treatment (B) . Samples from each treatment were analyzed by C1q binding ELISA. Columns represent the mean OD 450 value ± standard error from three samples.

    Techniques Used: Binding Assay, Purification, Incubation, SDS Page, Software, Expressing, Enzyme-linked Immunosorbent Assay

    Mouse immunization and serum titers. BALB/c mice (6 per group) were immunized twice two weeks apart subcutaneously with a dose that would deliver 8 μg ZE3 for each IgG fusion or with PBS as a control. Mouse serum samples were collected two weeks after the final dose (A) . Serially diluted mouse serum was analyzed for total IgG production by ELISA. The endpoint was taken as the reciprocal of the greatest dilution that gave an OD 450 reading at least twice the background. Two stars (**) indicates p
    Figure Legend Snippet: Mouse immunization and serum titers. BALB/c mice (6 per group) were immunized twice two weeks apart subcutaneously with a dose that would deliver 8 μg ZE3 for each IgG fusion or with PBS as a control. Mouse serum samples were collected two weeks after the final dose (A) . Serially diluted mouse serum was analyzed for total IgG production by ELISA. The endpoint was taken as the reciprocal of the greatest dilution that gave an OD 450 reading at least twice the background. Two stars (**) indicates p

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Various IgG fusion strategies enhance C1q binding (A) . Schematic representation of IgG fusion constructs used in this study. Fusion constructs are built on the mAb 6D8 human IgG1 backbone with the shown modifications. ZE3; the Zika envelope domain III containing amino acids K301 to T406; e, an epitope tag containing the “VYKLDISEA” 6D8 binding motif for RIC formation; e with lightning bolt, an epitope tag truncated to include only “YKLDIS” to reduce RIC formation; VH, the variable heavy domain from 6D8 which participates in binding the epitope tag; VL, the variably light domain from 6D8 which participates in binding the epitope tag; H, the heavy chain constant CH1 domain from 6D8; L, the light chain constant domain from 6D8; Fc, the heavy chain constant CH2 or CH3 domains from 6D8; Fc with lightning bolt, same as Fc but with E345R, E430G, and S440Y mutations to induce hexamer formation (B) . C1q binding ELISA of purified IgG fusion constructs made in plants that lack plant-specific β1,2-linked xylose and α1,3-linked fucose N-linked glycans. ELISA plates were coated with 15 μg/ml human C1q and incubated with 10 μg/ml each molecule, using 6D8 with no fusion as a negative control. Constructs were detected using polyclonal goat anti-human IgG-HRP. Mean OD 450 values from three samples are shown ± standard error with one star (*) indicating p
    Figure Legend Snippet: Various IgG fusion strategies enhance C1q binding (A) . Schematic representation of IgG fusion constructs used in this study. Fusion constructs are built on the mAb 6D8 human IgG1 backbone with the shown modifications. ZE3; the Zika envelope domain III containing amino acids K301 to T406; e, an epitope tag containing the “VYKLDISEA” 6D8 binding motif for RIC formation; e with lightning bolt, an epitope tag truncated to include only “YKLDIS” to reduce RIC formation; VH, the variable heavy domain from 6D8 which participates in binding the epitope tag; VL, the variably light domain from 6D8 which participates in binding the epitope tag; H, the heavy chain constant CH1 domain from 6D8; L, the light chain constant domain from 6D8; Fc, the heavy chain constant CH2 or CH3 domains from 6D8; Fc with lightning bolt, same as Fc but with E345R, E430G, and S440Y mutations to induce hexamer formation (B) . C1q binding ELISA of purified IgG fusion constructs made in plants that lack plant-specific β1,2-linked xylose and α1,3-linked fucose N-linked glycans. ELISA plates were coated with 15 μg/ml human C1q and incubated with 10 μg/ml each molecule, using 6D8 with no fusion as a negative control. Constructs were detected using polyclonal goat anti-human IgG-HRP. Mean OD 450 values from three samples are shown ± standard error with one star (*) indicating p

    Techniques Used: Binding Assay, Construct, Enzyme-linked Immunosorbent Assay, Purification, Incubation, Negative Control

    9) Product Images from "Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate"

    Article Title: Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiy377

    Study design. Three sequential Ebola virus (EBOV) challenge studies using Chinese-origin rhesus macaques were conducted to assess the postinfection efficacy of SAB-139 in early intervention ([EI] starting on 1 day [d] postinfection), delayed intervention ([DI] on 3 d postinfection) and late intervention ([LI] on 5 d postinfection). Collection of blood and cerebrospinal fluid samples was preplanned for each experiment, but collection differed by d based on group assignment and the experiment. (A) Experiment 1 evaluated 4 57 mg/kg intravenous (IV) doses of SAB-139/V3–V4 in an EI group (n = 6), an irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) control group (n = 6), or a saline control group (n = 2). (B) Experiment 2 evaluated three 150-mg/kg doses then one 125-mg/kg IV dose of SAB-139/V6–V8 in EI (n = 6) and DI (n = 6) groups or an irrelevant Tc-hIgG in the control group (n = 2). Surviving animals were back-challenged at d 77 after initial challenge to assess the development of an adaptive immune response postinfection and treatment. (C) Experiment 3 evaluated three 150-mg/kg doses then one 125-mg/kg dose of SAB-139/V6–V8 in a LI group (n = 6), an irrelevant Tc-hIgG in a control group (n = 4), or saline in a negative control group (n = 2). The * indicates that the fourth administration could be administered on d 11, 12, 13, or 14 postinfection based upon clinical and laboratory criteria. Abbreviation: EOS, end of study.
    Figure Legend Snippet: Study design. Three sequential Ebola virus (EBOV) challenge studies using Chinese-origin rhesus macaques were conducted to assess the postinfection efficacy of SAB-139 in early intervention ([EI] starting on 1 day [d] postinfection), delayed intervention ([DI] on 3 d postinfection) and late intervention ([LI] on 5 d postinfection). Collection of blood and cerebrospinal fluid samples was preplanned for each experiment, but collection differed by d based on group assignment and the experiment. (A) Experiment 1 evaluated 4 57 mg/kg intravenous (IV) doses of SAB-139/V3–V4 in an EI group (n = 6), an irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) control group (n = 6), or a saline control group (n = 2). (B) Experiment 2 evaluated three 150-mg/kg doses then one 125-mg/kg IV dose of SAB-139/V6–V8 in EI (n = 6) and DI (n = 6) groups or an irrelevant Tc-hIgG in the control group (n = 2). Surviving animals were back-challenged at d 77 after initial challenge to assess the development of an adaptive immune response postinfection and treatment. (C) Experiment 3 evaluated three 150-mg/kg doses then one 125-mg/kg dose of SAB-139/V6–V8 in a LI group (n = 6), an irrelevant Tc-hIgG in a control group (n = 4), or saline in a negative control group (n = 2). The * indicates that the fourth administration could be administered on d 11, 12, 13, or 14 postinfection based upon clinical and laboratory criteria. Abbreviation: EOS, end of study.

    Techniques Used: Negative Control

    Ebola virus (EBOV)-specific transchromosomic bovine (Tc-bovine)-derived human immunoglobulin G (hIgG) potently recruits multiple innate immune effector functions. (A and B) Induction of antibody-mediated monocyte phagocytosis (ADMP) (A) or antibody-mediated neutrophil phagocytosis (ADNP) (B) using antibodies from SAB-139/V3–V4 (blue), SAB-139/V6–V8 (purple), sera from Tc-bovines (2314 [black squares] and 2316 [white circles]), anti-EBOV monoclonal antibodies c13C6 (green) and KZ52 (orange), no antibody control (white), and irrelevant Tc IgG control (red). Uptake of EBOV glycoprotein (GP) onto coated fluorescein isothiocyanate (FITC) beads was determined by flow cytometry. A phagocytic score was determined using the percentage of FITC + cells and the mean fluorescent intensity (MFI) of the FITC + cells. The average phagocytic score of 2 replicates is shown. (C) The percentage of natural killer (NK) cells expressing CD107a protein and production of the cytokine, interferon (IFN)-γ, and the chemokine macrophage inflammatory protein (MIP)-1b was determined by flow cytometry (average of 2 replicates). (D) EBOV-specific IgG subclass MFI was determined via Luminex EBOV GP-coupled beads followed by detection with a subclass-specific secondary antibody (average of 2 replicates). (E) Correlation analyses using Spearman correlation coefficients between ADMP, ADNP, CD107a, IFNγ, MIP-1β, and EBOV GP-specific subclass titers (total IgG, IgG1–4) indicated a significant correlation between indicated function and/or titer, as marked by an asterisk. The r s values are represented by a heat map (range, −1 to 1).
    Figure Legend Snippet: Ebola virus (EBOV)-specific transchromosomic bovine (Tc-bovine)-derived human immunoglobulin G (hIgG) potently recruits multiple innate immune effector functions. (A and B) Induction of antibody-mediated monocyte phagocytosis (ADMP) (A) or antibody-mediated neutrophil phagocytosis (ADNP) (B) using antibodies from SAB-139/V3–V4 (blue), SAB-139/V6–V8 (purple), sera from Tc-bovines (2314 [black squares] and 2316 [white circles]), anti-EBOV monoclonal antibodies c13C6 (green) and KZ52 (orange), no antibody control (white), and irrelevant Tc IgG control (red). Uptake of EBOV glycoprotein (GP) onto coated fluorescein isothiocyanate (FITC) beads was determined by flow cytometry. A phagocytic score was determined using the percentage of FITC + cells and the mean fluorescent intensity (MFI) of the FITC + cells. The average phagocytic score of 2 replicates is shown. (C) The percentage of natural killer (NK) cells expressing CD107a protein and production of the cytokine, interferon (IFN)-γ, and the chemokine macrophage inflammatory protein (MIP)-1b was determined by flow cytometry (average of 2 replicates). (D) EBOV-specific IgG subclass MFI was determined via Luminex EBOV GP-coupled beads followed by detection with a subclass-specific secondary antibody (average of 2 replicates). (E) Correlation analyses using Spearman correlation coefficients between ADMP, ADNP, CD107a, IFNγ, MIP-1β, and EBOV GP-specific subclass titers (total IgG, IgG1–4) indicated a significant correlation between indicated function and/or titer, as marked by an asterisk. The r s values are represented by a heat map (range, −1 to 1).

    Techniques Used: Derivative Assay, Flow Cytometry, Cytometry, Expressing, Luminex

    SAB-139 can reduce or eliminate morbidity and mortality in rhesus macaques. (A) Experiment 1: Of the early-intervention (EI) nonhuman primates (NHPs) receiving SAB-139/V3–V4 intravenous (IV), 33% survived (2 of 6) compared with 0% (0 of 6) after administration irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) or normal saline (0 of 2) controls. Surviving animals had mild clinical signs but no detectable viremia (10 5  genome equivalents [GE]/mL limit of quantification [LOQ]). The NHPs had low anti-Ebola virus (EBOV) glycoprotein (GP) enzyme-linked immunosorbent assay (ELISA) titers after infusion (day [d] 3, 2908–7042 ELISA units [EU]/mL). One surviving NHP abruptly developed high anti-EBOV GP ELISA titers near end of study (EOS). (B) Experiment 2: Of EI and delayed intervention (DI) NHPs treated with SAB-139/V6–V8 IV, 100% survived (6 of 6 in both groups) compared with 0% of irrelevant Tc-hIgG controls (0 of 2). All SAB-treated NHPs were asymptomatic, but 2 EI animals had low sporadic viremia. All EI and DI NHPs had anti-EBOV GP ELISA titers after infusion that peaked at d 14 postinfection before falling at EOS. (C) Experiment 3: Of late intervention NHPs treated with SAB-139/V6–V8 IV, 0% survived (0 of 6) as did controls. All NHPs had viremia ranging from 10 8  to 10 10  GE/mL at d 5 postinfection. All animals died before or on d 9 postinfection. All NHPs had a low anti-EBOV GP ELISA titer at necropsy (1688–3629 EU/mL). Abbreviation: GGT, gamma-glutamyl transferase.
    Figure Legend Snippet: SAB-139 can reduce or eliminate morbidity and mortality in rhesus macaques. (A) Experiment 1: Of the early-intervention (EI) nonhuman primates (NHPs) receiving SAB-139/V3–V4 intravenous (IV), 33% survived (2 of 6) compared with 0% (0 of 6) after administration irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) or normal saline (0 of 2) controls. Surviving animals had mild clinical signs but no detectable viremia (10 5 genome equivalents [GE]/mL limit of quantification [LOQ]). The NHPs had low anti-Ebola virus (EBOV) glycoprotein (GP) enzyme-linked immunosorbent assay (ELISA) titers after infusion (day [d] 3, 2908–7042 ELISA units [EU]/mL). One surviving NHP abruptly developed high anti-EBOV GP ELISA titers near end of study (EOS). (B) Experiment 2: Of EI and delayed intervention (DI) NHPs treated with SAB-139/V6–V8 IV, 100% survived (6 of 6 in both groups) compared with 0% of irrelevant Tc-hIgG controls (0 of 2). All SAB-treated NHPs were asymptomatic, but 2 EI animals had low sporadic viremia. All EI and DI NHPs had anti-EBOV GP ELISA titers after infusion that peaked at d 14 postinfection before falling at EOS. (C) Experiment 3: Of late intervention NHPs treated with SAB-139/V6–V8 IV, 0% survived (0 of 6) as did controls. All NHPs had viremia ranging from 10 8 to 10 10 GE/mL at d 5 postinfection. All animals died before or on d 9 postinfection. All NHPs had a low anti-EBOV GP ELISA titer at necropsy (1688–3629 EU/mL). Abbreviation: GGT, gamma-glutamyl transferase.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    10) Product Images from "IgA binds to the AD‐2 epitope of glycoprotein B and neutralizes human cytomegalovirus"

    Article Title: IgA binds to the AD‐2 epitope of glycoprotein B and neutralizes human cytomegalovirus

    Journal: Immunology

    doi: 10.1111/imm.13286

    Scatterplot correlations of individual samples displaying IgG and IgA binding to both HCMV gB and the AD‐2 epitope. ELISA data displaying comparative IgG (A) and IgA (B) binding to gB and AD‐2 epitope of HCMV in each sample of human serum ( n = 24) and human saliva ( n = 24). Samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal and vertical dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples, respectively, were based on IgG gB‐negative samples ( n = 4). Correlation coefficients (Spearman's) and statistical significance for each set of determinations comparing serum or saliva IgG and IgA binding to gB and AD‐2 epitope are displayed within each graph
    Figure Legend Snippet: Scatterplot correlations of individual samples displaying IgG and IgA binding to both HCMV gB and the AD‐2 epitope. ELISA data displaying comparative IgG (A) and IgA (B) binding to gB and AD‐2 epitope of HCMV in each sample of human serum ( n = 24) and human saliva ( n = 24). Samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal and vertical dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples, respectively, were based on IgG gB‐negative samples ( n = 4). Correlation coefficients (Spearman's) and statistical significance for each set of determinations comparing serum or saliva IgG and IgA binding to gB and AD‐2 epitope are displayed within each graph

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    AD‐2 specific IgA mAbs neutralize HCMV in vitro. (A) Isotype‐matched mAbs 8F9 (IgG1 and IgA1) and QG1 (IgG1 and IgA1) binding to NS0 cells engineered to express the N‐terminal fragment of gB (NS0‐gBNT) as measured by flow cytometry. Grey filled histogram, secondary Ab alone: mAb – 2 µg/ml (black line), 1 µg/ml (dark grey line) and 0.2 µg/ml (light grey line). (B) Neutralization assays for HCMV infection of human foreskin fibroblasts (HFFs) represented as percentage of cells infected with high passage Merlin strain of HCMV. 8F9 IgG1, 8F9 IgA1, QG1 IgG1 and QG1 IgA1 were titrated with data shown as representative of 2 independently performed experiments. (C) Neutralization assay for HCMV (TB40/e) infection of human epithelial cells (ARPE‐19) represented as infection relative to no Ab control (assigned 1.0). 8F9 IgA1 and QG1 IgA1 were diluted 1:3 and assayed in triplicate. Statistical differences between the mean values of each mAb compared with no mAb for each concentration were obtained from the Mann–Whitney test (no asterisk is not significant; * P
    Figure Legend Snippet: AD‐2 specific IgA mAbs neutralize HCMV in vitro. (A) Isotype‐matched mAbs 8F9 (IgG1 and IgA1) and QG1 (IgG1 and IgA1) binding to NS0 cells engineered to express the N‐terminal fragment of gB (NS0‐gBNT) as measured by flow cytometry. Grey filled histogram, secondary Ab alone: mAb – 2 µg/ml (black line), 1 µg/ml (dark grey line) and 0.2 µg/ml (light grey line). (B) Neutralization assays for HCMV infection of human foreskin fibroblasts (HFFs) represented as percentage of cells infected with high passage Merlin strain of HCMV. 8F9 IgG1, 8F9 IgA1, QG1 IgG1 and QG1 IgA1 were titrated with data shown as representative of 2 independently performed experiments. (C) Neutralization assay for HCMV (TB40/e) infection of human epithelial cells (ARPE‐19) represented as infection relative to no Ab control (assigned 1.0). 8F9 IgA1 and QG1 IgA1 were diluted 1:3 and assayed in triplicate. Statistical differences between the mean values of each mAb compared with no mAb for each concentration were obtained from the Mann–Whitney test (no asterisk is not significant; * P

    Techniques Used: In Vitro, Binding Assay, Flow Cytometry, Neutralization, Infection, Concentration Assay, MANN-WHITNEY

    Human milk‐derived IgG and IgA bind HCMV gB and AD‐2. (A) ELISA to determine IgG and IgA binding to gB and AD‐2 epitope of HCMV in human breastmilk samples ( n = 14). Samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples were based on IgG gB‐negative samples ( n = 4). Statistical differences comparing the mean OD450 nm values of IgG and IgA binding to gB and AD‐2 epitope at each dilution in part A were obtained from the Mann–Whitney test (ns, not significant; ** P
    Figure Legend Snippet: Human milk‐derived IgG and IgA bind HCMV gB and AD‐2. (A) ELISA to determine IgG and IgA binding to gB and AD‐2 epitope of HCMV in human breastmilk samples ( n = 14). Samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples were based on IgG gB‐negative samples ( n = 4). Statistical differences comparing the mean OD450 nm values of IgG and IgA binding to gB and AD‐2 epitope at each dilution in part A were obtained from the Mann–Whitney test (ns, not significant; ** P

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

    Human IgG and IgA bind HCMV gB and AD‐2. ELISA to determine IgG and IgA binding to gB and AD‐2 epitope of HCMV in (A) human serum ( n = 24) and (B) human saliva ( n = 24). All samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples were based on IgG gB‐negative samples ( n = 4). Statistical differences comparing the mean OD450 nm values of IgG and IgA binding to gB and AD‐2 epitope at each dilution were obtained from the Mann–Whitney test (ns, not significant; **** P
    Figure Legend Snippet: Human IgG and IgA bind HCMV gB and AD‐2. ELISA to determine IgG and IgA binding to gB and AD‐2 epitope of HCMV in (A) human serum ( n = 24) and (B) human saliva ( n = 24). All samples were diluted as indicated, and data are represented as OD450 nm values of triplicate determinations of three independent experiments. Horizontal dotted lines representing positive/negative cut‐off values for negative gB and negative AD‐2 samples were based on IgG gB‐negative samples ( n = 4). Statistical differences comparing the mean OD450 nm values of IgG and IgA binding to gB and AD‐2 epitope at each dilution were obtained from the Mann–Whitney test (ns, not significant; **** P

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

    Recombinant antibodies expressed as human IgG and IgA bind gB and AD‐2 epitope of HCMV. (A) Expression of monoclonal recombinant antibodies QG1 IgG1, QG1 IgA1, 8F9 IgG1, 8F9 IgA1, FA9 IgG1 and FA9 IgA1 by transient transfection and ELISA of undiluted culture supernatants and binding to gB and AD‐2. +ve control, diluted human serum; –ve control, undiluted culture supernatant from untransfected cells; unrelated IgG and IgA are control human mAbs. (B) Binding and titration of concentration‐matched mAbs QG1 IgG1 (black bars), QG1 IgA1 (grey bars), 8F9 IgG1 (black bars), 8F9 IgA1 (grey bars) to gB and AD‐2 epitope of HCMV represented as OD values with human serum as positive control and unrelated IgG and unrelated IgA as negative control. Statistical differences between the mean OD values of IgG and IgA expression and for binding to gB and AD‐2 epitope were obtained from the Mann–Whitney test (** P
    Figure Legend Snippet: Recombinant antibodies expressed as human IgG and IgA bind gB and AD‐2 epitope of HCMV. (A) Expression of monoclonal recombinant antibodies QG1 IgG1, QG1 IgA1, 8F9 IgG1, 8F9 IgA1, FA9 IgG1 and FA9 IgA1 by transient transfection and ELISA of undiluted culture supernatants and binding to gB and AD‐2. +ve control, diluted human serum; –ve control, undiluted culture supernatant from untransfected cells; unrelated IgG and IgA are control human mAbs. (B) Binding and titration of concentration‐matched mAbs QG1 IgG1 (black bars), QG1 IgA1 (grey bars), 8F9 IgG1 (black bars), 8F9 IgA1 (grey bars) to gB and AD‐2 epitope of HCMV represented as OD values with human serum as positive control and unrelated IgG and unrelated IgA as negative control. Statistical differences between the mean OD values of IgG and IgA expression and for binding to gB and AD‐2 epitope were obtained from the Mann–Whitney test (** P

    Techniques Used: Recombinant, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Binding Assay, Titration, Concentration Assay, Positive Control, Negative Control, MANN-WHITNEY

    gB vaccination induces IgG and IgA responses to gB and AD‐2 in sero‐positive recipients. Serum samples obtained from the recombinant gB vaccination trial were analysed for IgG and IgA binding to gB and AD‐2 by ELISA. Individuals were stratified into (A) sero‐positive (IgG+to gB) and (B) sero‐negative (IgG‐ to gB). Statistical differences between the mean OD values of day 0 and 56 of vaccine recipients for IgG and IgA for binding to gB and AD‐2 epitope were obtained from the Mann–Whitney test (ns, not significant; * P
    Figure Legend Snippet: gB vaccination induces IgG and IgA responses to gB and AD‐2 in sero‐positive recipients. Serum samples obtained from the recombinant gB vaccination trial were analysed for IgG and IgA binding to gB and AD‐2 by ELISA. Individuals were stratified into (A) sero‐positive (IgG+to gB) and (B) sero‐negative (IgG‐ to gB). Statistical differences between the mean OD values of day 0 and 56 of vaccine recipients for IgG and IgA for binding to gB and AD‐2 epitope were obtained from the Mann–Whitney test (ns, not significant; * P

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

    11) Product Images from "IgG subclass distribution of the rheumatoid arthritis-specific autoantibodies to citrullinated fibrin"

    Article Title: IgG subclass distribution of the rheumatoid arthritis-specific autoantibodies to citrullinated fibrin

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/j.1365-2249.2004.02708.x

    Titration of the MoAbs to IgG subclasses. The curves correspond to the optimal dilution (see Methods) of the anti-IgG1, -IgG2, -IgG3 or -IgG4 MoAb, i.e. the dilution giving the same OD for the same concentration of the corresponding detected IgG subclass. Curves are the means of at least five duplicate determinations, performed in independent experiments. Bars indicate standard error. Titration curves obtained with the unlabelled NL16 anti-IgG1 MoAb (1 : 9000) followed by HRP-labelled antimouse IgG F(ab)′ 2 (1 : 2500), with the HRP-labelled HP6014 anti-IgG2 MoAb (1 : 2500), the HRP-labelled HP6050 anti-IgG3 MoAb (1 : 1500) and the HRP-labelled HP6025 anti-IgG4 MoAb (1 : 6000) are indicated by arrows. The thick full line represents the reference curve obtained with the HRP-labelled JDC10 anti-IgG MoAb (1 : 3000).
    Figure Legend Snippet: Titration of the MoAbs to IgG subclasses. The curves correspond to the optimal dilution (see Methods) of the anti-IgG1, -IgG2, -IgG3 or -IgG4 MoAb, i.e. the dilution giving the same OD for the same concentration of the corresponding detected IgG subclass. Curves are the means of at least five duplicate determinations, performed in independent experiments. Bars indicate standard error. Titration curves obtained with the unlabelled NL16 anti-IgG1 MoAb (1 : 9000) followed by HRP-labelled antimouse IgG F(ab)′ 2 (1 : 2500), with the HRP-labelled HP6014 anti-IgG2 MoAb (1 : 2500), the HRP-labelled HP6050 anti-IgG3 MoAb (1 : 1500) and the HRP-labelled HP6025 anti-IgG4 MoAb (1 : 6000) are indicated by arrows. The thick full line represents the reference curve obtained with the HRP-labelled JDC10 anti-IgG MoAb (1 : 3000).

    Techniques Used: Titration, Concentration Assay

    12) Product Images from "Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate"

    Article Title: Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiy377

    SAB-139 can reduce or eliminate morbidity and mortality in rhesus macaques. (A) Experiment 1: Of the early-intervention (EI) nonhuman primates (NHPs) receiving SAB-139/V3–V4 intravenous (IV), 33% survived (2 of 6) compared with 0% (0 of 6) after administration irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) or normal saline (0 of 2) controls. Surviving animals had mild clinical signs but no detectable viremia (10 5  genome equivalents [GE]/mL limit of quantification [LOQ]). The NHPs had low anti-Ebola virus (EBOV) glycoprotein (GP) enzyme-linked immunosorbent assay (ELISA) titers after infusion (day [d] 3, 2908–7042 ELISA units [EU]/mL). One surviving NHP abruptly developed high anti-EBOV GP ELISA titers near end of study (EOS). (B) Experiment 2: Of EI and delayed intervention (DI) NHPs treated with SAB-139/V6–V8 IV, 100% survived (6 of 6 in both groups) compared with 0% of irrelevant Tc-hIgG controls (0 of 2). All SAB-treated NHPs were asymptomatic, but 2 EI animals had low sporadic viremia. All EI and DI NHPs had anti-EBOV GP ELISA titers after infusion that peaked at d 14 postinfection before falling at EOS. (C) Experiment 3: Of late intervention NHPs treated with SAB-139/V6–V8 IV, 0% survived (0 of 6) as did controls. All NHPs had viremia ranging from 10 8  to 10 10  GE/mL at d 5 postinfection. All animals died before or on d 9 postinfection. All NHPs had a low anti-EBOV GP ELISA titer at necropsy (1688–3629 EU/mL). Abbreviation: GGT, gamma-glutamyl transferase.
    Figure Legend Snippet: SAB-139 can reduce or eliminate morbidity and mortality in rhesus macaques. (A) Experiment 1: Of the early-intervention (EI) nonhuman primates (NHPs) receiving SAB-139/V3–V4 intravenous (IV), 33% survived (2 of 6) compared with 0% (0 of 6) after administration irrelevant transchromosomic human immunoglobulin G (Tc-hIgG) or normal saline (0 of 2) controls. Surviving animals had mild clinical signs but no detectable viremia (10 5 genome equivalents [GE]/mL limit of quantification [LOQ]). The NHPs had low anti-Ebola virus (EBOV) glycoprotein (GP) enzyme-linked immunosorbent assay (ELISA) titers after infusion (day [d] 3, 2908–7042 ELISA units [EU]/mL). One surviving NHP abruptly developed high anti-EBOV GP ELISA titers near end of study (EOS). (B) Experiment 2: Of EI and delayed intervention (DI) NHPs treated with SAB-139/V6–V8 IV, 100% survived (6 of 6 in both groups) compared with 0% of irrelevant Tc-hIgG controls (0 of 2). All SAB-treated NHPs were asymptomatic, but 2 EI animals had low sporadic viremia. All EI and DI NHPs had anti-EBOV GP ELISA titers after infusion that peaked at d 14 postinfection before falling at EOS. (C) Experiment 3: Of late intervention NHPs treated with SAB-139/V6–V8 IV, 0% survived (0 of 6) as did controls. All NHPs had viremia ranging from 10 8 to 10 10 GE/mL at d 5 postinfection. All animals died before or on d 9 postinfection. All NHPs had a low anti-EBOV GP ELISA titer at necropsy (1688–3629 EU/mL). Abbreviation: GGT, gamma-glutamyl transferase.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    13) Product Images from "Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana"

    Article Title: Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana

    Journal: Plant biotechnology journal

    doi: 10.1111/j.1467-7652.2011.00593.x

    Anti-Ebola IgG responses in mice immunized with EIC. Groups of 7 mice were immunized subcutaneously with EIC, VRP-GP1, or PBS. The immunizations were given on days 0, 21, 42, and 63. The serum was collected on day 84 and assayed for anti-Ebola IgG by ELISA, using gamma-irradiated Ebola virus as the capture antigen. The data are presented as Geometric Mean Titer (GMT) ± the standard deviation for each group of mice.
    Figure Legend Snippet: Anti-Ebola IgG responses in mice immunized with EIC. Groups of 7 mice were immunized subcutaneously with EIC, VRP-GP1, or PBS. The immunizations were given on days 0, 21, 42, and 63. The serum was collected on day 84 and assayed for anti-Ebola IgG by ELISA, using gamma-irradiated Ebola virus as the capture antigen. The data are presented as Geometric Mean Titer (GMT) ± the standard deviation for each group of mice.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Irradiation, Standard Deviation

    Diagram illustrating the possible structure of the recombinant IgG and its assembly to form EIC. At left, the 6D8 H2-GP1 fusion protein (blue H2 chain, green GP1) is assembled with the light chain (red) to form a chimeric IgG-GP1. The 6D8 epitope on GP1 is shown as a red star. The epitope binding site at the top of the molecule can bind to the 6D8 epitope displayed on other chimeric molecules, which results is complex assembly (middle, with IgG component shown in purple). The complement component C1q can bind to the Fc region of IgG molecules that are bound to antigen (right).
    Figure Legend Snippet: Diagram illustrating the possible structure of the recombinant IgG and its assembly to form EIC. At left, the 6D8 H2-GP1 fusion protein (blue H2 chain, green GP1) is assembled with the light chain (red) to form a chimeric IgG-GP1. The 6D8 epitope on GP1 is shown as a red star. The epitope binding site at the top of the molecule can bind to the 6D8 epitope displayed on other chimeric molecules, which results is complex assembly (middle, with IgG component shown in purple). The complement component C1q can bind to the Fc region of IgG molecules that are bound to antigen (right).

    Techniques Used: Recombinant, Binding Assay

    Western blotting of EIC. a. Crude extract from N. benthamiana leaf agroinfiltrated with pBYH2GP1kdel, pBYK3R and pPS19. The 100 mg leaf sample was harvested 4 dpi and extracted with 500μl SDS sample buffer containing DTT, and 20μl was loaded on the gel. Western blot was probed with anti human IgG heavy chain. b-f. Purified EIC from leaves agroinfiltrated with pBYH2GP1kdelK3 and pPS19. b. Coomassie stained SDS-PAGE reducing gel; c. Reducing western blot detected with anti-human IgG heavy chain; d. Reducing western blot detected with anti human kappa chain; e. Reducing western blot detected with anti-linear epitope 6D8 mAb; f. Non-reducing western blot detected with anti-conformational epitope 13C6.
    Figure Legend Snippet: Western blotting of EIC. a. Crude extract from N. benthamiana leaf agroinfiltrated with pBYH2GP1kdel, pBYK3R and pPS19. The 100 mg leaf sample was harvested 4 dpi and extracted with 500μl SDS sample buffer containing DTT, and 20μl was loaded on the gel. Western blot was probed with anti human IgG heavy chain. b-f. Purified EIC from leaves agroinfiltrated with pBYH2GP1kdelK3 and pPS19. b. Coomassie stained SDS-PAGE reducing gel; c. Reducing western blot detected with anti-human IgG heavy chain; d. Reducing western blot detected with anti human kappa chain; e. Reducing western blot detected with anti-linear epitope 6D8 mAb; f. Non-reducing western blot detected with anti-conformational epitope 13C6.

    Techniques Used: Western Blot, Purification, Staining, SDS Page

    Expression of Ebola immune complex in N. benthamiana plants. a. Protein expression of Ebola immune complex compared among different constructs. N. benthamiana leaves were co-infiltrated with pBYH2GP1+pBYK3R+p19, pBYH2GP1kdel+pBYK3R+p19, and pBYRH2GP1kdelK3R+p19 at final OD600=0.25. The leaves were harvested 4 days after infiltration and extracted for ELISA to quantify IgG (Experimental Procedures) b. Protein expression level at different times after agroinfiltration using pBYRH2GP1kdK3 with pPSp19. The leaves were harvested on days 2, 4, 6, and 8 dpi. Data are means ± SD of samples from three independent infiltration experiments.
    Figure Legend Snippet: Expression of Ebola immune complex in N. benthamiana plants. a. Protein expression of Ebola immune complex compared among different constructs. N. benthamiana leaves were co-infiltrated with pBYH2GP1+pBYK3R+p19, pBYH2GP1kdel+pBYK3R+p19, and pBYRH2GP1kdelK3R+p19 at final OD600=0.25. The leaves were harvested 4 days after infiltration and extracted for ELISA to quantify IgG (Experimental Procedures) b. Protein expression level at different times after agroinfiltration using pBYRH2GP1kdK3 with pPSp19. The leaves were harvested on days 2, 4, 6, and 8 dpi. Data are means ± SD of samples from three independent infiltration experiments.

    Techniques Used: Expressing, Construct, Enzyme-linked Immunosorbent Assay

    Size measurement confirmed the complex formation. a. Hydrodynamic diameters of EIC or 6D8 mAb determined by dynamic light scattering using Zetasizer Nano-ZS instrument (Malvern Instruments, UK). The abscissa indicates the diameter in nm, and the ordinate indicates the relative number of molecules at that size, comparing human IgG, plant-expressed 6D8 mAb, and plant-expressed EIC. b. Size exclusion chromatography was used to determine the size of the immune complex. EIC was loaded onto BioSep SEC-S4000 (Phenomenex, USA) and eluted with PBS, pH7.3. The elution time in min is shown on the abscissa, and the A 280 was continuously monitored (colored curves). Bovine thyroglobulin (Sigma, USA), mouse IgG (SouthernBiotech, USA), and plant-expressed 6D8 mAb were used as protein markers.
    Figure Legend Snippet: Size measurement confirmed the complex formation. a. Hydrodynamic diameters of EIC or 6D8 mAb determined by dynamic light scattering using Zetasizer Nano-ZS instrument (Malvern Instruments, UK). The abscissa indicates the diameter in nm, and the ordinate indicates the relative number of molecules at that size, comparing human IgG, plant-expressed 6D8 mAb, and plant-expressed EIC. b. Size exclusion chromatography was used to determine the size of the immune complex. EIC was loaded onto BioSep SEC-S4000 (Phenomenex, USA) and eluted with PBS, pH7.3. The elution time in min is shown on the abscissa, and the A 280 was continuously monitored (colored curves). Bovine thyroglobulin (Sigma, USA), mouse IgG (SouthernBiotech, USA), and plant-expressed 6D8 mAb were used as protein markers.

    Techniques Used: Size-exclusion Chromatography

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    Incubation:

    Article Title: Loss of Humoral and Cellular Immunity to Invasive Nontyphoidal Salmonella during Current or Convalescent Plasmodium falciparum Infection in Malawian Children
    Article Snippet: Plates were washed with PBS containing 0.05% Tween 20 and blocked with 200 μl/well blocking buffer (PBS–1% bovine serum albumin [BSA]) for 1 h at 37°C. .. Test serum prepared at 1:20 in dilution buffer (PBS–0.05% Tween 20–1% BSA) was serially diluted 3-fold and incubated at 37°C for 1 h. After washing, 100 μl of 1:2,000 secondary goat anti-human IgG-AP antibodies (Southern Biotech) was added and incubated for 1 h at 37°C. .. Finally, after washing, 100 μl of SigmaFAST p-nitrophenyl phosphate substrate was added to each plate and the plate was read after 30 min using a BioTek ELx800 reader (BioTek Instruments, USA) at 405 nm.

    Article Title: Human Immunoglobulin G2 (IgG2) and IgG4, but Not IgG1 or IgG3, Protect Mice against Cryptococcus neoformans Infection ▿
    Article Snippet: .. Yeast cells were washed with PBS-1% BSA with 5 mM EDTA and then incubated in the same buffer for 30 min at 4°C first with 1:100 rat MAb to mouse C1q (Caltag Laboratories) and then with 1:100 rabbit F(ab′)2 anti-rat IgG-FITC cross-adsorbed against human IgG (Southern Biotechnology). .. After a final wash in PBS-1% BSA, yeast cells were analyzed by flow cytometry using a FACScan instrument (Becton Dickinson) equipped with a blue laser excitation of 15 mW at 488 nm.

    Expressing:

    Article Title: A Highly Expressing, Soluble, and Stable Plant-Made IgG Fusion Vaccine Strategy Enhances Antigen Immunogenicity in Mice Without Adjuvant
    Article Snippet: These data suggest that RIC solubility can be enhanced by reducing self-binding without a loss of C1q activation. .. Highly Variable Expression and Solubility of IgG Fusions in Plants Since a viable vaccine candidate must be soluble and highly expressing, we measured the soluble yield of fully assembled IgG fusions by employing an ELISA assay that first captured ZE3 and then detected human IgG. .. As a standard, purified and quantified HLZ was used.

    Solubility:

    Article Title: A Highly Expressing, Soluble, and Stable Plant-Made IgG Fusion Vaccine Strategy Enhances Antigen Immunogenicity in Mice Without Adjuvant
    Article Snippet: These data suggest that RIC solubility can be enhanced by reducing self-binding without a loss of C1q activation. .. Highly Variable Expression and Solubility of IgG Fusions in Plants Since a viable vaccine candidate must be soluble and highly expressing, we measured the soluble yield of fully assembled IgG fusions by employing an ELISA assay that first captured ZE3 and then detected human IgG. .. As a standard, purified and quantified HLZ was used.

    Enzyme-linked Immunosorbent Assay:

    Article Title: A Highly Expressing, Soluble, and Stable Plant-Made IgG Fusion Vaccine Strategy Enhances Antigen Immunogenicity in Mice Without Adjuvant
    Article Snippet: These data suggest that RIC solubility can be enhanced by reducing self-binding without a loss of C1q activation. .. Highly Variable Expression and Solubility of IgG Fusions in Plants Since a viable vaccine candidate must be soluble and highly expressing, we measured the soluble yield of fully assembled IgG fusions by employing an ELISA assay that first captured ZE3 and then detected human IgG. .. As a standard, purified and quantified HLZ was used.

    Article Title:
    Article Snippet: FuGENE6 transfection reagent was obtained from Roche Applied Science. .. Goat anti-human IgG and goat anti-human IgG-AP for the enzyme-linked immunosorbent assay were from Southern Biotech. .. Novex 4–12% gels and other electrophoresis reagents were from Invitrogen.

    Purification:

    Article Title: Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana
    Article Snippet: The diameter size was measured by Dynamic Light Scattering using Zetasizer Nano-ZS instrument (Malvern Instruments, UK). .. The purified plant-made Ebola immune complex, purified plant-made 6D8 mAb, and human IgG (Southern biotech, AL) were diluted in PBS to the concentration 0.1mg/ml and added into the disposable polystyrene cuvette for Zetasizer measurement. .. A sample volume of 20 μl was loaded onto BioSep SEC-S4000 column, 600X7.8 mm (Phenomenex, Torrance, CA).

    Concentration Assay:

    Article Title: Expression of an immunogenic Ebola immune complex in Nicotiana benthamiana
    Article Snippet: The diameter size was measured by Dynamic Light Scattering using Zetasizer Nano-ZS instrument (Malvern Instruments, UK). .. The purified plant-made Ebola immune complex, purified plant-made 6D8 mAb, and human IgG (Southern biotech, AL) were diluted in PBS to the concentration 0.1mg/ml and added into the disposable polystyrene cuvette for Zetasizer measurement. .. A sample volume of 20 μl was loaded onto BioSep SEC-S4000 column, 600X7.8 mm (Phenomenex, Torrance, CA).

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  • 98
    SouthernBiotech igg
    Binding of membrane-permeable TAT-Runx1.d190 protein to the murine c-Myc locus. (A) Schematic of TAT-Runx1.d190 fusion protein. The Runx1.d190 protein lacks the TAT peptide, but has N-terminal and C-terminal polyhistidine tags. (B) Differential affinity of <t>polyclonal</t> antisera recognizing the N-terminus of distal Runx1 for Runx1.d190 and TAT-Runx1.d190 proteins. Histidine-tagged Runx1.d190 (lanes 2 and 3), or TAT-Runx1.d190 protein (lanes 4 and 6) adhered to nickel beads, or nickel beads alone (lanes 1 and 5) were fixed in 1% formaldeyde to simulate ChIP conditions and incubated with control Pre-immune sera (lanes 2 and 4) or anti-distal Runx1 (α-Runx1) antisera (lanes 1, 3, 5 and 6). The amount of pre-immune or anti-distal Runx1 immunoglobin associated with fixed Runx1.d190 or TAT-Runx1.d190 proteins is shown on a representative immunoblot probed with anti-immunoglobin (Anti-Ig). The amount of Runx1.d190 or TAT-Runx1.d190 proteins immobilized on the beads prior to fixation is shown on a representative immunoblot probed with anti-polyhistidine (Anti-histidine). N =3. (C) ChIP analysis of chromatin from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with preimmune sera (Pre-immune) or anti-distal Runx1 (α-Runx1). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. A representative of three independent experiments is shown. (D) The PCR product yields from chromatin from TAT peptide- or TAT-Runx1.d190-treated murine splenocytes immunoprecipitated with anti-distal Runx1, relative to input, are graphed. The PCR primers used are indicated on the x-axis. P-values derived from a two-tailed t test indicating statistical significance are shown above the brackets. (E) ChIP analysis of chromatin prepared from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with control <t>IgG</t> (IgG) or anti-polyhistidine (α-Histidine). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. N =3.
    Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg/product/SouthernBiotech
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    93
    SouthernBiotech pe conjugated mouse anti human total igg
    HIV infection is associated with modified M. tuberculosis -specific total <t>IgG</t> titers. Relative levels of IgG specific to PPD (A), LAM (B), Ag85A/B (C), ESAT6/CFP10 (D), HIV gp120 (E), influenza HA (F), PPSV23 (G), and tetanus toxoid (H) present in the plasma of the study population and negative controls were determined via Luminex. The MFI for each individual is graphed. The gray dotted line is the median of the control group. Within each violin plot, the black solid line is the median and the black dashed lines show the interquartile range. Kruskal-Wallis with Dunn’s multiple-comparison test was used. Adjusted P values are as follows: *, P
    Pe Conjugated Mouse Anti Human Total Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    SouthernBiotech igg isotype specific antibodies
    Serum <t>IgM,</t> <t>IgG,</t> and IgA levels over time in the patient.
    Igg Isotype Specific Antibodies, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg isotype specific antibodies/product/SouthernBiotech
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    97
    SouthernBiotech anti mouse igg subclass hrp specific antibodies
    Production and characterization of αDEC-NS1 and αDCIR2-NS1 mAbs. Approximately 1 µg of each mAb was separated under reduced (A) and non-reduced (B) conditions. Both gels were stained with Coomassie Blue dye. The heavy (HC) and light (LC) chains of the following antibodies are shown: 1) αDEC, 2) αDCIR2, 3) αDEC-NS1, 4) αDCIR2-NS1. (C) Western blotting using anti-mouse <t>IgG</t> and anti-mouse kappa chain conjugated to <t>HRP</t> to verify the recognition of both heavy and light chains of each antibody. The numbers indicate the same order as in (A). (D) Western blotting on a non-reduced gel using a mouse serum raised against the dimeric form of NS1 followed by protein A conjugated to HRP. Columns 1–4, same as (A), column 5 contains 1 µg of NS1 produced in bacteria (Di_NS1: dimeric NS1 and Mo_NS1: monomeric NS1). Specific reaction was obtained only in columns containing NS1. MW, molecular weight marker in kDa.
    Anti Mouse Igg Subclass Hrp Specific Antibodies, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Binding of membrane-permeable TAT-Runx1.d190 protein to the murine c-Myc locus. (A) Schematic of TAT-Runx1.d190 fusion protein. The Runx1.d190 protein lacks the TAT peptide, but has N-terminal and C-terminal polyhistidine tags. (B) Differential affinity of polyclonal antisera recognizing the N-terminus of distal Runx1 for Runx1.d190 and TAT-Runx1.d190 proteins. Histidine-tagged Runx1.d190 (lanes 2 and 3), or TAT-Runx1.d190 protein (lanes 4 and 6) adhered to nickel beads, or nickel beads alone (lanes 1 and 5) were fixed in 1% formaldeyde to simulate ChIP conditions and incubated with control Pre-immune sera (lanes 2 and 4) or anti-distal Runx1 (α-Runx1) antisera (lanes 1, 3, 5 and 6). The amount of pre-immune or anti-distal Runx1 immunoglobin associated with fixed Runx1.d190 or TAT-Runx1.d190 proteins is shown on a representative immunoblot probed with anti-immunoglobin (Anti-Ig). The amount of Runx1.d190 or TAT-Runx1.d190 proteins immobilized on the beads prior to fixation is shown on a representative immunoblot probed with anti-polyhistidine (Anti-histidine). N =3. (C) ChIP analysis of chromatin from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with preimmune sera (Pre-immune) or anti-distal Runx1 (α-Runx1). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. A representative of three independent experiments is shown. (D) The PCR product yields from chromatin from TAT peptide- or TAT-Runx1.d190-treated murine splenocytes immunoprecipitated with anti-distal Runx1, relative to input, are graphed. The PCR primers used are indicated on the x-axis. P-values derived from a two-tailed t test indicating statistical significance are shown above the brackets. (E) ChIP analysis of chromatin prepared from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with control IgG (IgG) or anti-polyhistidine (α-Histidine). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. N =3.

    Journal: PLoS ONE

    Article Title: Runx Transcription Factors Repress Human and Murine c-Myc Expression in a DNA-Binding and C-Terminally Dependent Manner

    doi: 10.1371/journal.pone.0069083

    Figure Lengend Snippet: Binding of membrane-permeable TAT-Runx1.d190 protein to the murine c-Myc locus. (A) Schematic of TAT-Runx1.d190 fusion protein. The Runx1.d190 protein lacks the TAT peptide, but has N-terminal and C-terminal polyhistidine tags. (B) Differential affinity of polyclonal antisera recognizing the N-terminus of distal Runx1 for Runx1.d190 and TAT-Runx1.d190 proteins. Histidine-tagged Runx1.d190 (lanes 2 and 3), or TAT-Runx1.d190 protein (lanes 4 and 6) adhered to nickel beads, or nickel beads alone (lanes 1 and 5) were fixed in 1% formaldeyde to simulate ChIP conditions and incubated with control Pre-immune sera (lanes 2 and 4) or anti-distal Runx1 (α-Runx1) antisera (lanes 1, 3, 5 and 6). The amount of pre-immune or anti-distal Runx1 immunoglobin associated with fixed Runx1.d190 or TAT-Runx1.d190 proteins is shown on a representative immunoblot probed with anti-immunoglobin (Anti-Ig). The amount of Runx1.d190 or TAT-Runx1.d190 proteins immobilized on the beads prior to fixation is shown on a representative immunoblot probed with anti-polyhistidine (Anti-histidine). N =3. (C) ChIP analysis of chromatin from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with preimmune sera (Pre-immune) or anti-distal Runx1 (α-Runx1). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. A representative of three independent experiments is shown. (D) The PCR product yields from chromatin from TAT peptide- or TAT-Runx1.d190-treated murine splenocytes immunoprecipitated with anti-distal Runx1, relative to input, are graphed. The PCR primers used are indicated on the x-axis. P-values derived from a two-tailed t test indicating statistical significance are shown above the brackets. (E) ChIP analysis of chromatin prepared from murine splenocytes treated with 0.5 µM TAT peptide or TAT-Runx1.d190 protein and immunoprecipitated with control IgG (IgG) or anti-polyhistidine (α-Histidine). PCR was carried out with primer pairs amplifying consensus Runx-binding sites at -4.3 (ii), -5.4 (iii), and -7.6 (iv) kb upstream of the murine c-Myc transcriptional start site. N =3.

    Article Snippet: Chromatin was immunoprecipitated with the control antibody rabbit pre-immune sera, rabbit polyclonal murine anti-distal Runx1 [ ], IgG (Southern Biotech, Birmingham, AL) or a His-probe (AD 1.1.10) antibody (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Binding Assay, Chromatin Immunoprecipitation, Incubation, Immunoprecipitation, Polymerase Chain Reaction, Derivative Assay, Two Tailed Test

    HIV infection is associated with modified M. tuberculosis -specific total IgG titers. Relative levels of IgG specific to PPD (A), LAM (B), Ag85A/B (C), ESAT6/CFP10 (D), HIV gp120 (E), influenza HA (F), PPSV23 (G), and tetanus toxoid (H) present in the plasma of the study population and negative controls were determined via Luminex. The MFI for each individual is graphed. The gray dotted line is the median of the control group. Within each violin plot, the black solid line is the median and the black dashed lines show the interquartile range. Kruskal-Wallis with Dunn’s multiple-comparison test was used. Adjusted P values are as follows: *, P

    Journal: mSphere

    Article Title: HIV Is Associated with Modified Humoral Immune Responses in the Setting of HIV/TB Coinfection

    doi: 10.1128/mSphere.00104-20

    Figure Lengend Snippet: HIV infection is associated with modified M. tuberculosis -specific total IgG titers. Relative levels of IgG specific to PPD (A), LAM (B), Ag85A/B (C), ESAT6/CFP10 (D), HIV gp120 (E), influenza HA (F), PPSV23 (G), and tetanus toxoid (H) present in the plasma of the study population and negative controls were determined via Luminex. The MFI for each individual is graphed. The gray dotted line is the median of the control group. Within each violin plot, the black solid line is the median and the black dashed lines show the interquartile range. Kruskal-Wallis with Dunn’s multiple-comparison test was used. Adjusted P values are as follows: *, P

    Article Snippet: The plate was then washed 6 times with assay buffer, and 40 μl of PE-conjugated mouse anti-human total IgG, IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, or IgM (Southern Biotech) at 1.3 μg/ml was added and incubated with shaking at 800 rpm at RT for 1 h. The plate was then washed 6 times with sheath fluid (Luminex Corporation) and resuspended in sheath fluid in a final volume of 60 μl.

    Techniques: Infection, Modification, Laser Capture Microdissection, Luminex

    Bulk immunoglobulin levels are increased during HIV and M. tuberculosis infection. Relative levels of bulk IgG1 (A), IgG2 (B), IgG3 (C), IgG4 (D), IgA (E), and IgM (F) present in the plasma of the study population and negative controls were determined via Luminex. The median fluorescence intensity (MFI) for each individual is graphed. The gray dotted lines is the median of the control group. Within each violin plot, the black solid line is the median and the black dashed lines show the interquartile range. Kruskal-Wallis with Dunn’s multiple-comparison test was used. Adjusted P values are as follows: *, P

    Journal: mSphere

    Article Title: HIV Is Associated with Modified Humoral Immune Responses in the Setting of HIV/TB Coinfection

    doi: 10.1128/mSphere.00104-20

    Figure Lengend Snippet: Bulk immunoglobulin levels are increased during HIV and M. tuberculosis infection. Relative levels of bulk IgG1 (A), IgG2 (B), IgG3 (C), IgG4 (D), IgA (E), and IgM (F) present in the plasma of the study population and negative controls were determined via Luminex. The median fluorescence intensity (MFI) for each individual is graphed. The gray dotted lines is the median of the control group. Within each violin plot, the black solid line is the median and the black dashed lines show the interquartile range. Kruskal-Wallis with Dunn’s multiple-comparison test was used. Adjusted P values are as follows: *, P

    Article Snippet: The plate was then washed 6 times with assay buffer, and 40 μl of PE-conjugated mouse anti-human total IgG, IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, or IgM (Southern Biotech) at 1.3 μg/ml was added and incubated with shaking at 800 rpm at RT for 1 h. The plate was then washed 6 times with sheath fluid (Luminex Corporation) and resuspended in sheath fluid in a final volume of 60 μl.

    Techniques: Infection, Luminex, Fluorescence

    Antigen-specific IgM and IgG4 titers correlate with CD4 + T cell counts. (A) Spearman correlations across all HIV+ individuals comparing CD4 + T cell counts and viral loads to Z-scored antigen-specific isotype and subclass titers. Statistically significant correlations ( P

    Journal: mSphere

    Article Title: HIV Is Associated with Modified Humoral Immune Responses in the Setting of HIV/TB Coinfection

    doi: 10.1128/mSphere.00104-20

    Figure Lengend Snippet: Antigen-specific IgM and IgG4 titers correlate with CD4 + T cell counts. (A) Spearman correlations across all HIV+ individuals comparing CD4 + T cell counts and viral loads to Z-scored antigen-specific isotype and subclass titers. Statistically significant correlations ( P

    Article Snippet: The plate was then washed 6 times with assay buffer, and 40 μl of PE-conjugated mouse anti-human total IgG, IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, or IgM (Southern Biotech) at 1.3 μg/ml was added and incubated with shaking at 800 rpm at RT for 1 h. The plate was then washed 6 times with sheath fluid (Luminex Corporation) and resuspended in sheath fluid in a final volume of 60 μl.

    Techniques:

    Serum IgM, IgG, and IgA levels over time in the patient.

    Journal:

    Article Title: CD20 deficiency in humans results in impaired T cell-independent antibody responses

    doi: 10.1172/JCI40231

    Figure Lengend Snippet: Serum IgM, IgG, and IgA levels over time in the patient.

    Article Snippet: The plates were washed, incubated with alkaline phosphatase–conjugated polyclonal goat anti-mouse IgM and IgG isotype-specific antibodies (Southern Biotechnology Associates) for 1 hour at room temperature, and developed using p -nitrophenyl phosphate substrate (Sigma-Aldrich).

    Techniques:

    Production and characterization of αDEC-NS1 and αDCIR2-NS1 mAbs. Approximately 1 µg of each mAb was separated under reduced (A) and non-reduced (B) conditions. Both gels were stained with Coomassie Blue dye. The heavy (HC) and light (LC) chains of the following antibodies are shown: 1) αDEC, 2) αDCIR2, 3) αDEC-NS1, 4) αDCIR2-NS1. (C) Western blotting using anti-mouse IgG and anti-mouse kappa chain conjugated to HRP to verify the recognition of both heavy and light chains of each antibody. The numbers indicate the same order as in (A). (D) Western blotting on a non-reduced gel using a mouse serum raised against the dimeric form of NS1 followed by protein A conjugated to HRP. Columns 1–4, same as (A), column 5 contains 1 µg of NS1 produced in bacteria (Di_NS1: dimeric NS1 and Mo_NS1: monomeric NS1). Specific reaction was obtained only in columns containing NS1. MW, molecular weight marker in kDa.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Targeting the Non-structural Protein 1 from Dengue Virus to a Dendritic Cell Population Confers Protective Immunity to Lethal Virus Challenge

    doi: 10.1371/journal.pntd.0002330

    Figure Lengend Snippet: Production and characterization of αDEC-NS1 and αDCIR2-NS1 mAbs. Approximately 1 µg of each mAb was separated under reduced (A) and non-reduced (B) conditions. Both gels were stained with Coomassie Blue dye. The heavy (HC) and light (LC) chains of the following antibodies are shown: 1) αDEC, 2) αDCIR2, 3) αDEC-NS1, 4) αDCIR2-NS1. (C) Western blotting using anti-mouse IgG and anti-mouse kappa chain conjugated to HRP to verify the recognition of both heavy and light chains of each antibody. The numbers indicate the same order as in (A). (D) Western blotting on a non-reduced gel using a mouse serum raised against the dimeric form of NS1 followed by protein A conjugated to HRP. Columns 1–4, same as (A), column 5 contains 1 µg of NS1 produced in bacteria (Di_NS1: dimeric NS1 and Mo_NS1: monomeric NS1). Specific reaction was obtained only in columns containing NS1. MW, molecular weight marker in kDa.

    Article Snippet: The secondary antibodies used were goat anti-mouse IgG Fc-specific-HRP (1∶5,000; Jackson ImmunoResearch Laboratories) or anti-mouse IgG subclass-HRP specific antibodies (1∶3,000; SouthernBiotech).

    Techniques: Staining, Western Blot, Produced, Molecular Weight, Marker