idez protease igg specific  (New England Biolabs)


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    New England Biolabs idez protease igg specific
    <t>IdeZ</t> rescues AAV8 and AAV9 liver transduction in passively immunized mice and cynomolgus macaques with preexisting NAbs. ( A ) Experimental timeline of <t>IgG,</t> IdeZ, and AAV8- or AAV9-Luc injections. Sac., sacrifice followed by tissue harvest. Mice were injected IP with pooled human IgG. The same mice were injected IV 24 hours later with PBS or recombinant GST-IdeZ (2.5 mg/kg). AAV8-Luc or AAV9-Luc was injected 72 hours post–IdeZ treatment at a dose of 1 × 10 13 vg/kg. Luciferase transgene expression levels were analyzed 4 weeks postinjection in the liver: AAV8 ( B and C ) and AAV9 ( D and E ). Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. Biodistribution of AAV8- and AAV9-Luc vector genomes in the liver: AAV8 ( F and G ) and AAV9 ( H and I ). Vector genome copy numbers per cell were calculated by normalizing Luc copy numbers to copies of the lamin B2 housekeeping gene and represented as log vg/cell. Each dot represents a technical duplicate from a single animal, and the dash represents the mean value. (F, female; M, male.) ( J ) Schematic demonstrating experimental timeline of IdeZ and AAV9-Luc injections in nonhuman primates (NHPs). AAV9-seropositive NHP M16558 ( n = 1) was administered IdeZ (0.5 mg/kg) via IV bolus injection on day 0. AAV9-Luc was administered via IV bolus injection 72 hours post–IdeZ injection at a dose of 5 × 10 12 vg/kg. ( K ) NHP serum samples were analyzed by SDS-PAGE under reducing conditions and probed with Fc-specific antibodies. ( L ) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the livers of NHPs. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents a single experiment of an individual liver lobe from a single animal. ( M ) Biodistribution of AAV9-Luc vector genomes in the livers of NHPs. Vector genome copy numbers per nanogram of total extracted DNA were calculated and represented as log vg/ng DNA. Each dot represents a technical duplicate experiment of individual liver slices from a single animal, and the dash represents the mean value. Significance was determined by 1-way ANOVA with Tukey’s posttest. * P
    Idez Protease Igg Specific, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/idez protease igg specific/product/New England Biolabs
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    idez protease igg specific - by Bioz Stars, 2022-12
    94/100 stars

    Images

    1) Product Images from "Rescuing AAV gene transfer from neutralizing antibodies with an IgG-degrading enzyme"

    Article Title: Rescuing AAV gene transfer from neutralizing antibodies with an IgG-degrading enzyme

    Journal: JCI Insight

    doi: 10.1172/jci.insight.139881

    IdeZ rescues AAV8 and AAV9 liver transduction in passively immunized mice and cynomolgus macaques with preexisting NAbs. ( A ) Experimental timeline of IgG, IdeZ, and AAV8- or AAV9-Luc injections. Sac., sacrifice followed by tissue harvest. Mice were injected IP with pooled human IgG. The same mice were injected IV 24 hours later with PBS or recombinant GST-IdeZ (2.5 mg/kg). AAV8-Luc or AAV9-Luc was injected 72 hours post–IdeZ treatment at a dose of 1 × 10 13 vg/kg. Luciferase transgene expression levels were analyzed 4 weeks postinjection in the liver: AAV8 ( B and C ) and AAV9 ( D and E ). Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. Biodistribution of AAV8- and AAV9-Luc vector genomes in the liver: AAV8 ( F and G ) and AAV9 ( H and I ). Vector genome copy numbers per cell were calculated by normalizing Luc copy numbers to copies of the lamin B2 housekeeping gene and represented as log vg/cell. Each dot represents a technical duplicate from a single animal, and the dash represents the mean value. (F, female; M, male.) ( J ) Schematic demonstrating experimental timeline of IdeZ and AAV9-Luc injections in nonhuman primates (NHPs). AAV9-seropositive NHP M16558 ( n = 1) was administered IdeZ (0.5 mg/kg) via IV bolus injection on day 0. AAV9-Luc was administered via IV bolus injection 72 hours post–IdeZ injection at a dose of 5 × 10 12 vg/kg. ( K ) NHP serum samples were analyzed by SDS-PAGE under reducing conditions and probed with Fc-specific antibodies. ( L ) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the livers of NHPs. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents a single experiment of an individual liver lobe from a single animal. ( M ) Biodistribution of AAV9-Luc vector genomes in the livers of NHPs. Vector genome copy numbers per nanogram of total extracted DNA were calculated and represented as log vg/ng DNA. Each dot represents a technical duplicate experiment of individual liver slices from a single animal, and the dash represents the mean value. Significance was determined by 1-way ANOVA with Tukey’s posttest. * P
    Figure Legend Snippet: IdeZ rescues AAV8 and AAV9 liver transduction in passively immunized mice and cynomolgus macaques with preexisting NAbs. ( A ) Experimental timeline of IgG, IdeZ, and AAV8- or AAV9-Luc injections. Sac., sacrifice followed by tissue harvest. Mice were injected IP with pooled human IgG. The same mice were injected IV 24 hours later with PBS or recombinant GST-IdeZ (2.5 mg/kg). AAV8-Luc or AAV9-Luc was injected 72 hours post–IdeZ treatment at a dose of 1 × 10 13 vg/kg. Luciferase transgene expression levels were analyzed 4 weeks postinjection in the liver: AAV8 ( B and C ) and AAV9 ( D and E ). Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. Biodistribution of AAV8- and AAV9-Luc vector genomes in the liver: AAV8 ( F and G ) and AAV9 ( H and I ). Vector genome copy numbers per cell were calculated by normalizing Luc copy numbers to copies of the lamin B2 housekeeping gene and represented as log vg/cell. Each dot represents a technical duplicate from a single animal, and the dash represents the mean value. (F, female; M, male.) ( J ) Schematic demonstrating experimental timeline of IdeZ and AAV9-Luc injections in nonhuman primates (NHPs). AAV9-seropositive NHP M16558 ( n = 1) was administered IdeZ (0.5 mg/kg) via IV bolus injection on day 0. AAV9-Luc was administered via IV bolus injection 72 hours post–IdeZ injection at a dose of 5 × 10 12 vg/kg. ( K ) NHP serum samples were analyzed by SDS-PAGE under reducing conditions and probed with Fc-specific antibodies. ( L ) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the livers of NHPs. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents a single experiment of an individual liver lobe from a single animal. ( M ) Biodistribution of AAV9-Luc vector genomes in the livers of NHPs. Vector genome copy numbers per nanogram of total extracted DNA were calculated and represented as log vg/ng DNA. Each dot represents a technical duplicate experiment of individual liver slices from a single animal, and the dash represents the mean value. Significance was determined by 1-way ANOVA with Tukey’s posttest. * P

    Techniques Used: Transduction, Mouse Assay, Injection, Recombinant, Luciferase, Expressing, Protein Concentration, Plasmid Preparation, SDS Page

    IdeZ cleaves serum antibodies from multiple species. ( A ) Schematic outlining IdeZ cleavage of IgG below the hinge region yielding multiple F(ab′) 2 and Fc fragments after reduction. ( B ) Serum samples from mouse, dog, primate, and human untreated (-) or treated (+) with recombinant IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. * indicates digested Fc/2 and Fd′ fragments. ( C ) Pooled human IgG untreated (-) or treated (+) with recombinant glutathione- S -transferase–IdeZ (GST-IdeZ) or commercial standard IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. IgG was cleaved by GST-IdeZ and IdeZ into multiple fragments as indicated. ( D ) Experimental timeline of in vivo GST-IdeZ dose optimization experiment. Mice were injected with pooled human IgG followed 24 hours later with no injection or injection with 3 different doses of GST-IdeZ. Blood serum samples were collected 72 hours post GST-IdeZ. Sac., sacrifice followed by tissue harvest. ( E ) Mice were injected IP first with pooled human IgG, followed by IV injection with PBS 24 hours later (-) or recombinant GST-IdeZ (1 mg/kg) (+). Blood samples were taken 72 hours after injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. IgG was probed with Fab- and Fc-specific antibodies. ( F ) Mice were injected IP first with pooled human IgG, followed by IV injection with PBS 24 hours later (-) or recombinant GST-IdeZ at 3 doses (0.25 mg/kg, 1 mg/kg, and 2.5 mg/kg) (+). Blood samples were taken 72 hours after injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. Human IgG was probed with an Fc-specific antibody. See complete unedited blots in the supplemental material.
    Figure Legend Snippet: IdeZ cleaves serum antibodies from multiple species. ( A ) Schematic outlining IdeZ cleavage of IgG below the hinge region yielding multiple F(ab′) 2 and Fc fragments after reduction. ( B ) Serum samples from mouse, dog, primate, and human untreated (-) or treated (+) with recombinant IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. * indicates digested Fc/2 and Fd′ fragments. ( C ) Pooled human IgG untreated (-) or treated (+) with recombinant glutathione- S -transferase–IdeZ (GST-IdeZ) or commercial standard IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. IgG was cleaved by GST-IdeZ and IdeZ into multiple fragments as indicated. ( D ) Experimental timeline of in vivo GST-IdeZ dose optimization experiment. Mice were injected with pooled human IgG followed 24 hours later with no injection or injection with 3 different doses of GST-IdeZ. Blood serum samples were collected 72 hours post GST-IdeZ. Sac., sacrifice followed by tissue harvest. ( E ) Mice were injected IP first with pooled human IgG, followed by IV injection with PBS 24 hours later (-) or recombinant GST-IdeZ (1 mg/kg) (+). Blood samples were taken 72 hours after injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. IgG was probed with Fab- and Fc-specific antibodies. ( F ) Mice were injected IP first with pooled human IgG, followed by IV injection with PBS 24 hours later (-) or recombinant GST-IdeZ at 3 doses (0.25 mg/kg, 1 mg/kg, and 2.5 mg/kg) (+). Blood samples were taken 72 hours after injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. Human IgG was probed with an Fc-specific antibody. See complete unedited blots in the supplemental material.

    Techniques Used: Recombinant, SDS Page, Staining, In Vivo, Mouse Assay, Injection, IV Injection

    Impact of IdeZ treatment on AAV9 cardiac transduction in passively immunized mice. ( A ) Experimental timeline of pooled human IgG, IdeZ, and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. ( B and D ) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the heart. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents the average of a technical duplicate from a single animal. ( C and E ) Biodistribution of AAV9-Luc vector genomes in the heart. Vector genome copy numbers per cell were calculated based on normalization to copies of the lamin B2 housekeeping gene. Each dot represents the average of a technical duplicate from a single animal. Significance was determined by 1-way ANOVA with Tukey’s posttest. * P
    Figure Legend Snippet: Impact of IdeZ treatment on AAV9 cardiac transduction in passively immunized mice. ( A ) Experimental timeline of pooled human IgG, IdeZ, and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. ( B and D ) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the heart. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents the average of a technical duplicate from a single animal. ( C and E ) Biodistribution of AAV9-Luc vector genomes in the heart. Vector genome copy numbers per cell were calculated based on normalization to copies of the lamin B2 housekeeping gene. Each dot represents the average of a technical duplicate from a single animal. Significance was determined by 1-way ANOVA with Tukey’s posttest. * P

    Techniques Used: Transduction, Mouse Assay, Derivative Assay, Luciferase, Expressing, Protein Concentration, Plasmid Preparation

    2) Product Images from "Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme"

    Article Title: Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme

    Journal: bioRxiv

    doi: 10.1101/2020.05.12.092122

    Impact of IdeZ treatment on AAV9 cardiac transduction in passively immunized mice. A , Experimental timeline of pooled human IgG, IdeZ and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. B,D , Luciferase transgene expression levels were analyzed 4 weeks post-injection in the heart. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. C,E , Biodistribution of AAV9 Luc vector genomes in the heart. Vector genome copy numbers per cell were calculated based on normalization to copies of the Lamin B2 housekeeping gene. Each dot represents the average of a technical duplicate from a single animal. Female (F), Male (M). Significance was determined one-way ANOVA with Tukey’s post-test. * p
    Figure Legend Snippet: Impact of IdeZ treatment on AAV9 cardiac transduction in passively immunized mice. A , Experimental timeline of pooled human IgG, IdeZ and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. B,D , Luciferase transgene expression levels were analyzed 4 weeks post-injection in the heart. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. C,E , Biodistribution of AAV9 Luc vector genomes in the heart. Vector genome copy numbers per cell were calculated based on normalization to copies of the Lamin B2 housekeeping gene. Each dot represents the average of a technical duplicate from a single animal. Female (F), Male (M). Significance was determined one-way ANOVA with Tukey’s post-test. * p

    Techniques Used: Transduction, Mouse Assay, Derivative Assay, Luciferase, Expressing, Injection, Protein Concentration, Plasmid Preparation

    IdeZ cleaves serum antibodies from multiple species. A , Schematic outlining IdeZ cleavage of IgG below the hinge region yielding multiple F(ab’) 2 and Fc fragments after reduction. B , Serum samples from mouse, dog, primate and human untreated (-) or treated (+) with recombinant IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. C , Pooled human IgG untreated (-) or treated (+) with recombinant GST-IdeZ or commercial standard IdeZ (NEB) and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. IgG was cleaved by GST-IdeZ and IdeZ into multiple fragments as indicated. D , Mice were injected intraperitoneally first with pooled human IgG, following which they were injected intravenously 24 hours later with PBS (-) or recombinant GST-IdeZ (1 mg/kg) (+). Blood samples were taken 72 hours post injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. IgG was probed with Fab (top panel) and Fc (bottom panel) specific antibodies. E , Experimental timeline of in vivo GST-IdeZ dose optimization experiment. Mice were injected with pooled human IgG followed 24 hrs later with no injection or injection with 3 different doses of GST-IdeZ. Blood serum samples were collected 72 hours post GST-IdeZ. Sac., sacrifice followed by tissue harvest. Serum samples of PBS control ( F ), 0.25 mg/kg ( G ), 1 mg/kg ( H ), and 2.5 mg/kg ( I ) GST-IdeZ injected mice were analyzed by SDS-PAGE under reducing conditions and probed with human IgG specific antibodies to analyze IgG cleavage.
    Figure Legend Snippet: IdeZ cleaves serum antibodies from multiple species. A , Schematic outlining IdeZ cleavage of IgG below the hinge region yielding multiple F(ab’) 2 and Fc fragments after reduction. B , Serum samples from mouse, dog, primate and human untreated (-) or treated (+) with recombinant IdeZ and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. C , Pooled human IgG untreated (-) or treated (+) with recombinant GST-IdeZ or commercial standard IdeZ (NEB) and analyzed by SDS-PAGE under reducing conditions. Gels were then stained with Coomassie blue. IgG was cleaved by GST-IdeZ and IdeZ into multiple fragments as indicated. D , Mice were injected intraperitoneally first with pooled human IgG, following which they were injected intravenously 24 hours later with PBS (-) or recombinant GST-IdeZ (1 mg/kg) (+). Blood samples were taken 72 hours post injection and analyzed by SDS-PAGE under reducing conditions with immunoblotting. IgG was probed with Fab (top panel) and Fc (bottom panel) specific antibodies. E , Experimental timeline of in vivo GST-IdeZ dose optimization experiment. Mice were injected with pooled human IgG followed 24 hrs later with no injection or injection with 3 different doses of GST-IdeZ. Blood serum samples were collected 72 hours post GST-IdeZ. Sac., sacrifice followed by tissue harvest. Serum samples of PBS control ( F ), 0.25 mg/kg ( G ), 1 mg/kg ( H ), and 2.5 mg/kg ( I ) GST-IdeZ injected mice were analyzed by SDS-PAGE under reducing conditions and probed with human IgG specific antibodies to analyze IgG cleavage.

    Techniques Used: Recombinant, SDS Page, Staining, Mouse Assay, Injection, In Vivo

    IdeZ rescues AAV8 and AAV9 liver transduction in passively immunized mice and cynomolgus macaques. A , Experimental timeline of IgG, IdeZ and AAV8 or AAV9-Luc injections. Sac., sacrifice followed by tissue harvest. Mice were injected intraperitoneally with pooled human IgG. The same mice were injected intravenously 24 hours later with PBS or recombinant GST-IdeZ (2.5 mg/kg). AAV8-Luc or AAV9-Luc was injected 72 hours post IdeZ at a dose of 1 × 10 13 vg/kg. Luciferase transgene expression levels were analyzed 4 weeks post-injection in the liver; AAV8 ( B,C ); AAV9 ( D,E ). Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. Biodistribution of AAV8 and AAV9 Luc vector genomes in the liver; AAV8 ( F,G ); AAV9 ( H,I ). Vector genome copy numbers per cell were calculated by normalizing Luc copy numbers to copies of the Lamin B2 housekeeping gene and represented as log vg/cell. Each dot represents a technical duplicate from a single animal, and the dash represents the mean value. (F=female, M=male). J , Schematic demonstrating experimental timeline of IdeZ and AAV9-Luc injections in NHPs. AAV9 seropositive NHP M16558 (n=1) was administered IdeZ (0.5 mg/kg) via intravenous bolus injection on Day 0. AAV9-Luc was administered via intravenous bolus injection 72 hrs post-IdeZ injection at a dose of 5 × 10 12 vg/kg. K , NHP serum samples were analyzed by SDS-PAGE under reducing conditions and probed with Fc specific antibodies. L , Luciferase transgene expression levels were analyzed 4 weeks post-injection in the liver of NHPs. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (Log RLU/g tissue). Each dot represents a single experiment of an individual liver lobe from a single animal. M , Biodistribution of AAV9 Luc vector genomes in the liver of NHPs. Vector genome copy numbers per ng of total extracted DNA were calculated and represented as log vg/ng DNA. Each dot represents a technical duplicate experiment of individual liver slices from a single animal and the dash represents the mean value. Significance was determined by one-way ANOVA with Tukey’s post-test. * p
    Figure Legend Snippet: IdeZ rescues AAV8 and AAV9 liver transduction in passively immunized mice and cynomolgus macaques. A , Experimental timeline of IgG, IdeZ and AAV8 or AAV9-Luc injections. Sac., sacrifice followed by tissue harvest. Mice were injected intraperitoneally with pooled human IgG. The same mice were injected intravenously 24 hours later with PBS or recombinant GST-IdeZ (2.5 mg/kg). AAV8-Luc or AAV9-Luc was injected 72 hours post IdeZ at a dose of 1 × 10 13 vg/kg. Luciferase transgene expression levels were analyzed 4 weeks post-injection in the liver; AAV8 ( B,C ); AAV9 ( D,E ). Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (log RLU/g tissue). Each dot represents the average of a technical duplicate from a single animal. Biodistribution of AAV8 and AAV9 Luc vector genomes in the liver; AAV8 ( F,G ); AAV9 ( H,I ). Vector genome copy numbers per cell were calculated by normalizing Luc copy numbers to copies of the Lamin B2 housekeeping gene and represented as log vg/cell. Each dot represents a technical duplicate from a single animal, and the dash represents the mean value. (F=female, M=male). J , Schematic demonstrating experimental timeline of IdeZ and AAV9-Luc injections in NHPs. AAV9 seropositive NHP M16558 (n=1) was administered IdeZ (0.5 mg/kg) via intravenous bolus injection on Day 0. AAV9-Luc was administered via intravenous bolus injection 72 hrs post-IdeZ injection at a dose of 5 × 10 12 vg/kg. K , NHP serum samples were analyzed by SDS-PAGE under reducing conditions and probed with Fc specific antibodies. L , Luciferase transgene expression levels were analyzed 4 weeks post-injection in the liver of NHPs. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue (Log RLU/g tissue). Each dot represents a single experiment of an individual liver lobe from a single animal. M , Biodistribution of AAV9 Luc vector genomes in the liver of NHPs. Vector genome copy numbers per ng of total extracted DNA were calculated and represented as log vg/ng DNA. Each dot represents a technical duplicate experiment of individual liver slices from a single animal and the dash represents the mean value. Significance was determined by one-way ANOVA with Tukey’s post-test. * p

    Techniques Used: Transduction, Mouse Assay, Injection, Recombinant, Luciferase, Expressing, Protein Concentration, Plasmid Preparation, SDS Page

    3) Product Images from "A genetic, genomic, and computational resource for exploring neural circuit function"

    Article Title: A genetic, genomic, and computational resource for exploring neural circuit function

    Journal: bioRxiv

    doi: 10.1101/385476

    Tandem-affinity purification of INTACT nuclei (TAPIN) enables neuronal genomics. A. Cell type-specific drivers enable expression of the UNC84-2XGFP nuclear tag (green) in specific populations of cells. Both the targeted cell type and driver are indicated in the lower left and right corner, respectively. B. Following nuclei harvest, two rounds of magnetic bead capture serially purify target nuclei. After the first round of protein A bead capture, bacterial protease IdeZ cleaves the anti-GFP antibody in the flexible hinge region, allowing a second round of bead capture with protein G, which recognizes the F(ab’) 2 region. Protein G, unlike Protein A, can bind both the Fc and F(ab’) 2 regions of an immunoglobulin. C. Two capture rounds reduce the level of non-specific background (grey bars, mock IgG control) while maintaining the cDNA yield from the captured target nuclei (green bars). Bars represent the mean of two replicates (shown as points). D. RNA-seq libraries created with more nuclei yield more cDNA (circles). TAPIN libraries had lower non-specific background than INTACT (blue vs orange triangles). E. Libraries with more cDNA detect more genes. F. Libraries with more cDNA have more reproducible transcript abundances. G. Previously identified markers of lamina monopolar and inner photoreceptor neurons ( Tan et al., 2015 ) are enriched in the expected cells.
    Figure Legend Snippet: Tandem-affinity purification of INTACT nuclei (TAPIN) enables neuronal genomics. A. Cell type-specific drivers enable expression of the UNC84-2XGFP nuclear tag (green) in specific populations of cells. Both the targeted cell type and driver are indicated in the lower left and right corner, respectively. B. Following nuclei harvest, two rounds of magnetic bead capture serially purify target nuclei. After the first round of protein A bead capture, bacterial protease IdeZ cleaves the anti-GFP antibody in the flexible hinge region, allowing a second round of bead capture with protein G, which recognizes the F(ab’) 2 region. Protein G, unlike Protein A, can bind both the Fc and F(ab’) 2 regions of an immunoglobulin. C. Two capture rounds reduce the level of non-specific background (grey bars, mock IgG control) while maintaining the cDNA yield from the captured target nuclei (green bars). Bars represent the mean of two replicates (shown as points). D. RNA-seq libraries created with more nuclei yield more cDNA (circles). TAPIN libraries had lower non-specific background than INTACT (blue vs orange triangles). E. Libraries with more cDNA detect more genes. F. Libraries with more cDNA have more reproducible transcript abundances. G. Previously identified markers of lamina monopolar and inner photoreceptor neurons ( Tan et al., 2015 ) are enriched in the expected cells.

    Techniques Used: Affinity Purification, Expressing, RNA Sequencing Assay

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    New England Biolabs idez protease igg specific
    Idez Protease Igg Specific, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/idez protease igg specific/product/New England Biolabs
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    idez protease igg specific - by Bioz Stars, 2022-12
    94/100 stars
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