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lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated  (Bioss)


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    Bioss lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated
    Lat (Tyr200) Polyclonal Antibody, Alexa Fluor 488 Conjugated, supplied by Bioss, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated/product/Bioss
    Average 91 stars, based on 2 article reviews
    lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated - by Bioz Stars, 2026-02
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    lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated  (Bioss)
    91
    Bioss lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated
    Lat (Tyr200) Polyclonal Antibody, Alexa Fluor 488 Conjugated, supplied by Bioss, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lat (tyr200) polyclonal antibody, alexa fluor 488 conjugated/product/Bioss
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    normal mouse igg2a af488  (Bioss)
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    Bioss normal mouse igg2a af488
    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Normal Mouse Igg2a Af488, supplied by Bioss, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lat tyr200  (Bioss)
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    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Lat Tyr200, supplied by Bioss, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bs 10128r a647  (Bioss)
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    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Bs 10128r A647, supplied by Bioss, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Polyclonal Catalog No, supplied by Bioss, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    af647 p lat tyr200  (Bioss)
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    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Af647 P Lat Tyr200, supplied by Bioss, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lat (tyr200) polyclonal antibody, alexa fluor 647 conjugated  (Bioss)
    90
    Bioss lat (tyr200) polyclonal antibody, alexa fluor 647 conjugated
    IRAP-deficient mice show less severe IgE- and <t>IgG-induced</t> anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.
    Lat (Tyr200) Polyclonal Antibody, Alexa Fluor 647 Conjugated, supplied by Bioss, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lat (tyr200) polyclonal antibody, alexa fluor 647 conjugated/product/Bioss
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    IRAP-deficient mice show less severe IgE- and IgG-induced anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.

    Journal: Frontiers in Immunology

    Article Title: Insulin-regulated aminopeptidase contributes to setting the intensity of FcR-mediated inflammation

    doi: 10.3389/fimmu.2022.1029759

    Figure Lengend Snippet: IRAP-deficient mice show less severe IgE- and IgG-induced anaphylactic reactions and experimental arthritis. (A) 24h after sensitization with anti-DNP IgE wild-type ( WT) and IRAP-deficient (IRAP KO ) mice were challenged with antigen (DNP-HSA) to induce passive systemic anaphylaxis (PSA). The drop in body temperature was evaluated. Data presented are the mean ± s.e.m. with 9 mice (pooled from 3 experiments). (B) Released serum MCPT-1 chymase collected at the end of temperature measurements was evaluated in the two groups of mice as well as in unsensitized control mice. (C) WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were immunized with rabbit IgG and 7 days later mice were challenged with rabbit IgG to induce active systemic anaphylaxis (ASA). The drop in body temperature was evaluated. Data are presented as mean ± s.e.m. with 6 mice (pooled from 2 experiments). (D) ASA was monitored by evaluating plasma platelet counts collected at the end of temperature measurements in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (E) Serum MCPT-1 chymase collected at the end of temperature measurements was measured in the two groups of mice as well as in non-immunized control mice. Data are the mean ± s.e.m. (F) WT and IRAP KO or WT FcγRIIA Tg and FcγRIIA Tg IRAP KO mice were injected with an anti-collagen type II Ab cocktail (day 0) followed by injection of LPS (day 4). Photographs shows representative hematoxylin/eosin (HE) staining of ankle sections as well as the macroscopic appearance of hind legs for each genotype at day 8. (G) Arthritis development was monitored by measuring paw thickness starting 5 days after injection of the Ab cocktail. (H) Arthritis scores were also evaluated according to the provided scoring system (Chondrex). Data are the mean ± s.e.m. from 4 to 7 mice/group. Statistical analysis was done using the two-way ANOVA followed by Sidak’s post-hoc test (A, C, G, H) or the unpaired Student’s t test (B, D, E) . *: P < 0.05; **: P < 0.01; *** P < 0.001; ns, not significant.

    Article Snippet: Abs used: human Syk (Tyr525 +Tyr526) Antibody (Bioss #bs-3434R-A488) (recognizing Tyr519 + Tyr520 in mouse sequence), Rabbit mAb IgG isotype control Alexa 488 (Cellsignaling 2975S), Syk (SantaCruz sc-1240 AF488), normal mouse IgG2a AF488 (SantaCruz sc-3891), LAT Tyr200 (Bioss bs-10128R-A488) (recognizing Tyr185 in mouse sequence), Rabbit IgG isotype control AF488 (Bioss bs-0295P-A488), LAT (SantaCruz sc-373706 AF790), Normal Mouse IgG2b (SantaCruz sc-516630 AF790), p-p38 Tyr182 (SantaCruz sc-166182 AF647), Normal mouse IgG2a AF647 (santacruz sc-24637), p38 (Santacruz sc-7972 AF405), Normal mouse IgG1 AF405 (Santacruz sc-45069), SHP-1 Signalway Antibody C32280-AF647), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S), Alexa Fluor ® 647 Anti-SHP1 (phospho Ser591) (Bioss # Bs-5578R), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S).

    Techniques: Injection, Staining

    Diminished phosphorylation response of signaling effectors in FcεRI-stimulated BMMCs and IgG stimulated neutrophils and monocytes in the context of IRAP-deficiency. (A–C) WT and IRAP-deficient (IRAP KO ) BMMCs were sensitized with anti-DNP IgE for 24 hours before stimulating them with specific antigen (DNP-HSA). After indicated time points, stimulation was arrested by adding fixation and permeabilization buffer. Phosflow analysis of signaling effectors (left panel) was assessed using anti-pSyk Y519/520 (A) , anti-pLAT Y200 (B) and anti-pp38 Y182 (C) . Total levels of proteins in non-stimulated cells were also analyzed using anti-Syk, anti-LAT and anti p38 (right panels). Phosphorylation levels were determined as the ratio of the gMFI of samples divided by the gMFI of the FMO of the respective sample and represent the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. (D, E) Phosflow analysis of Syk phosphorylation at Y 519/520 was also determined on ex vivo IgG-stimulated neutrophils and monocytes during ASA. Cells were accessed directly from the blood 5 min after initiation of ASA. The gating strategies for analysis of BMMCs, neutrophils and monocytes and representative examples of Ab staining are shown, respectively, in ( <xref ref-type= Supplementary Figures 2B, C ). Data shown are analyzed as in (A–C) and presented as the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. *: P < 0.05; **: P < 0.01; *** P < 0.001. ns, not significant. " width="100%" height="100%">

    Journal: Frontiers in Immunology

    Article Title: Insulin-regulated aminopeptidase contributes to setting the intensity of FcR-mediated inflammation

    doi: 10.3389/fimmu.2022.1029759

    Figure Lengend Snippet: Diminished phosphorylation response of signaling effectors in FcεRI-stimulated BMMCs and IgG stimulated neutrophils and monocytes in the context of IRAP-deficiency. (A–C) WT and IRAP-deficient (IRAP KO ) BMMCs were sensitized with anti-DNP IgE for 24 hours before stimulating them with specific antigen (DNP-HSA). After indicated time points, stimulation was arrested by adding fixation and permeabilization buffer. Phosflow analysis of signaling effectors (left panel) was assessed using anti-pSyk Y519/520 (A) , anti-pLAT Y200 (B) and anti-pp38 Y182 (C) . Total levels of proteins in non-stimulated cells were also analyzed using anti-Syk, anti-LAT and anti p38 (right panels). Phosphorylation levels were determined as the ratio of the gMFI of samples divided by the gMFI of the FMO of the respective sample and represent the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. (D, E) Phosflow analysis of Syk phosphorylation at Y 519/520 was also determined on ex vivo IgG-stimulated neutrophils and monocytes during ASA. Cells were accessed directly from the blood 5 min after initiation of ASA. The gating strategies for analysis of BMMCs, neutrophils and monocytes and representative examples of Ab staining are shown, respectively, in ( Supplementary Figures 2B, C ). Data shown are analyzed as in (A–C) and presented as the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. *: P < 0.05; **: P < 0.01; *** P < 0.001. ns, not significant.

    Article Snippet: Abs used: human Syk (Tyr525 +Tyr526) Antibody (Bioss #bs-3434R-A488) (recognizing Tyr519 + Tyr520 in mouse sequence), Rabbit mAb IgG isotype control Alexa 488 (Cellsignaling 2975S), Syk (SantaCruz sc-1240 AF488), normal mouse IgG2a AF488 (SantaCruz sc-3891), LAT Tyr200 (Bioss bs-10128R-A488) (recognizing Tyr185 in mouse sequence), Rabbit IgG isotype control AF488 (Bioss bs-0295P-A488), LAT (SantaCruz sc-373706 AF790), Normal Mouse IgG2b (SantaCruz sc-516630 AF790), p-p38 Tyr182 (SantaCruz sc-166182 AF647), Normal mouse IgG2a AF647 (santacruz sc-24637), p38 (Santacruz sc-7972 AF405), Normal mouse IgG1 AF405 (Santacruz sc-45069), SHP-1 Signalway Antibody C32280-AF647), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S), Alexa Fluor ® 647 Anti-SHP1 (phospho Ser591) (Bioss # Bs-5578R), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S).

    Techniques: Ex Vivo, Staining

    IRAP-deficient cells show less SHP1-inactivating phosphorylation on Ser591. (A) Anti-DNP IgE sensitized WT and IRAP-deficient (IRAP KO ) BMMCs were stimulated with specific antigen (30 ng/mL of DNP-HSA) for indicated time points and levels of the SHP1 S591 phosphorylation response were determined at indicated time points by phosflow analysis using anti-SHP1 S591 (left panel). Total levels of proteins of resting cells were analyzed using anti-SHP1 (right panel). (B, C) SHP1 phosphorylation on Ser591 was also determined on IgG-stimulated neutrophils (B) and monocytes (C) during ASA analyzed ex vivo using phosflow analysis. Cells were accessed directly from the blood 5 min after initiation of ASA. The gating strategies for analysis of BMMCs, neutrophils and monocytes and representative examples of Ab staining are shown, respectively in ( <xref ref-type= Supplementary Figures 2B, C ). Phosphorylation levels in (A–C) were determined as the ratio of the gMFI of samples divided by the gMFI of the Isotype of the respective sample and represent the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. *: P < 0.05; **: P < 0.01; ns, not significant. " width="100%" height="100%">

    Journal: Frontiers in Immunology

    Article Title: Insulin-regulated aminopeptidase contributes to setting the intensity of FcR-mediated inflammation

    doi: 10.3389/fimmu.2022.1029759

    Figure Lengend Snippet: IRAP-deficient cells show less SHP1-inactivating phosphorylation on Ser591. (A) Anti-DNP IgE sensitized WT and IRAP-deficient (IRAP KO ) BMMCs were stimulated with specific antigen (30 ng/mL of DNP-HSA) for indicated time points and levels of the SHP1 S591 phosphorylation response were determined at indicated time points by phosflow analysis using anti-SHP1 S591 (left panel). Total levels of proteins of resting cells were analyzed using anti-SHP1 (right panel). (B, C) SHP1 phosphorylation on Ser591 was also determined on IgG-stimulated neutrophils (B) and monocytes (C) during ASA analyzed ex vivo using phosflow analysis. Cells were accessed directly from the blood 5 min after initiation of ASA. The gating strategies for analysis of BMMCs, neutrophils and monocytes and representative examples of Ab staining are shown, respectively in ( Supplementary Figures 2B, C ). Phosphorylation levels in (A–C) were determined as the ratio of the gMFI of samples divided by the gMFI of the Isotype of the respective sample and represent the mean ± s.e.m of indicated experiments. Statistical analysis was done using a Student’s test. *: P < 0.05; **: P < 0.01; ns, not significant.

    Article Snippet: Abs used: human Syk (Tyr525 +Tyr526) Antibody (Bioss #bs-3434R-A488) (recognizing Tyr519 + Tyr520 in mouse sequence), Rabbit mAb IgG isotype control Alexa 488 (Cellsignaling 2975S), Syk (SantaCruz sc-1240 AF488), normal mouse IgG2a AF488 (SantaCruz sc-3891), LAT Tyr200 (Bioss bs-10128R-A488) (recognizing Tyr185 in mouse sequence), Rabbit IgG isotype control AF488 (Bioss bs-0295P-A488), LAT (SantaCruz sc-373706 AF790), Normal Mouse IgG2b (SantaCruz sc-516630 AF790), p-p38 Tyr182 (SantaCruz sc-166182 AF647), Normal mouse IgG2a AF647 (santacruz sc-24637), p38 (Santacruz sc-7972 AF405), Normal mouse IgG1 AF405 (Santacruz sc-45069), SHP-1 Signalway Antibody C32280-AF647), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S), Alexa Fluor ® 647 Anti-SHP1 (phospho Ser591) (Bioss # Bs-5578R), Rabbit mAb IgG XP ® Isotype Ctrl Alexa Fluor ® 647 Conjugate (Cell Signaling #2985S).

    Techniques: Ex Vivo, Staining