Structured Review

Agilent technologies rabbit polyclonal ab
Anti-hsp90 antibodies increase NadA Δ351–405 monocyte stimulation in a polymixin B insensitive-way. A) Cells were treated with NadA Δ351–405 in the absence (black circles) or in the presence (open circles) of purified rabbit <t>polyclonal</t> antibodies directed to the COOH terminal domain of hsp90. Open squares refers to cells incubated with NadA Δ351–405 in the presence of purified rabbit polyclonal antibodies from non immunized animals. The indicated cytokines/chemokines were analyzed in the extracellular medium by BioPlex suspension arrays. Data are the mean from a representative experiment out of four run in triplicate. Bars are +/− SE. Asterisk indicates signals significantly different (p
Rabbit Polyclonal Ab, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "The Soluble Recombinant Neisseria meningitidis Adhesin NadAΔ351–405 Stimulates Human Monocytes by Binding to Extracellular Hsp90"

Article Title: The Soluble Recombinant Neisseria meningitidis Adhesin NadAΔ351–405 Stimulates Human Monocytes by Binding to Extracellular Hsp90

Journal: PLoS ONE

doi: 10.1371/journal.pone.0025089

Anti-hsp90 antibodies increase NadA Δ351–405 monocyte stimulation in a polymixin B insensitive-way. A) Cells were treated with NadA Δ351–405 in the absence (black circles) or in the presence (open circles) of purified rabbit polyclonal antibodies directed to the COOH terminal domain of hsp90. Open squares refers to cells incubated with NadA Δ351–405 in the presence of purified rabbit polyclonal antibodies from non immunized animals. The indicated cytokines/chemokines were analyzed in the extracellular medium by BioPlex suspension arrays. Data are the mean from a representative experiment out of four run in triplicate. Bars are +/− SE. Asterisk indicates signals significantly different (p
Figure Legend Snippet: Anti-hsp90 antibodies increase NadA Δ351–405 monocyte stimulation in a polymixin B insensitive-way. A) Cells were treated with NadA Δ351–405 in the absence (black circles) or in the presence (open circles) of purified rabbit polyclonal antibodies directed to the COOH terminal domain of hsp90. Open squares refers to cells incubated with NadA Δ351–405 in the presence of purified rabbit polyclonal antibodies from non immunized animals. The indicated cytokines/chemokines were analyzed in the extracellular medium by BioPlex suspension arrays. Data are the mean from a representative experiment out of four run in triplicate. Bars are +/− SE. Asterisk indicates signals significantly different (p

Techniques Used: Purification, Incubation

Related Articles

Fluorescence:

Article Title: Involvement of the Tyro3 receptor and its intracellular partner Fyn signaling in Schwann cell myelination
Article Snippet: .. AntibodiesThe following antibodies were purchased: rabbit monoclonal anti-Tyro3 (5585S), rabbit monoclonal anti–neuronal marker GAP43 (8945S), rabbit monoclonal anti–68-kDa neurofilament NF68 (2835S), rabbit polyclonal anti-(pY527)Src (corresponding to pY531 in Fyn), and rabbit monoclonal anti–apoptotic marker active, cleaved caspase-3 (9579S) from Cell Signaling Technology (Danvers, MA); mouse monoclonal neuronal axon marker anti–βIII tubulin (MAB1195) from R & D Systems (Minneapolis, MN); mouse monoclonal anti-Fyn (05-436) and mouse monoclonal anti-(pS473)Akt (05-1003) from Merck-Millipore (Billerica, MA); rabbit polyclonal anti-Fyn (HPA023887) from Atlas Antibodies (Stockholm, Sweden); rabbit polyclonal anti–proliferating cell nuclear marker Ki67 (A0047) from Agilent Technology (Santa Clara, CA); mouse monoclonal anti-(pY416)Src (corresponding to pY420 in Fyn; sc-81521), mouse monoclonal anti-Akt1 (sc-5298), rabbit polyclonal anti-Oct6 (also called Scip; sc-133865), and rabbit polyclonal anti-Krox20 (also called Egr2; sc-20690) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit polyclonal anti-Tyro3 (ab109231) and mouse monoclonal anti-S100β (ab52642) from Abcam (Cambridge, United Kingdom); mouse monoclonal anti-MBP (SMI-94R-100) and monoclonal anti-GFAP (SMI-22R-100) from Covance (Princeton, NJ); anti-MPZ (also called P0) from MBL (Nagoya, Japan; rabbit polyclonal PD046) and Abnova (Taipei, Taiwan; goat polyclonal, PAB7332); mouse monoclonal anti–pan-sodium channel (K58/35) from Sigma-Aldrich (St. Louis, MO); mouse monoclonal anti-Rac1 (610652), mouse monoclonal anti-Cdc42 (610928), and mouse monoclonal anti–β-actin (612656) from Becton Dickinson (Franklin Lakes, NJ); mouse monoclonal anti–V5-epitope-tag (04434-94) from Nacalai Tesque (Kyoto, Japan); peroxidase-conjugated secondary antibodies from GE Healthcare (Fairfield, CT) and Nacalai Tesque; and fluorescence-labeled secondary antibodies from Life Technologies (Carlsbad, CA) and Wako Chemicals (Osaka, Japan). ..

Blocking Assay:

Article Title: von Willebrand factor self-association on platelet GpIb? under hydrodynamic shear: effect on shear-induced platelet activation
Article Snippet: .. Anti-VWF Abs include rabbit polyclonal Ab (Dako North America), nonfunction blocking mAb AVW-1 against the C-terminus (GTI Diagnostics), mAb AVW-3 against VWF-A1 domain (GTI Diagnostics), mAb SZ-123 against VWF-A3 domain (gift from Dr C. Ruan, Suzhou University, Suzhou, China) and 242.6 against VWF propeptide (gift from Dr R. Montgomery, BloodCenter of Wisconsin, Milwaukee, WI). .. Anti-GpIbα/CD42b function blocking mAbs are AP-1 (GTI Diagnostics), AK2 (Millipore), and HIP1 (BD Biosciences).

Activation Assay:

Article Title: Rheumatoid Factors Induce Signaling from B Cells, Leading to Epstein-Barr Virus and B-Cell Activation
Article Snippet: .. The Abs used for EBV activation and B-cell activation were a rabbit polyclonal Ab to human IgG (γ-chain-specific) (Dako, Copenhagen, Denmark), F(ab′)2 fragment of mouse monoclonal Ab (MAb) to the Fc fragment of human IgG (Jackson ImmunoResearch, West Grove, Pa.), and the F(ab′)2 fragment of mouse MAb to the Fab fragment of human IgG (Jackson ImmunoResearch). .. Other Abs included mouse MAb to phosphotyrosine (Cell Signaling, Beverly, Mass.), rabbit polyclonal Abs to phospho-Syk and phospho-ERK (Cell Signaling), a mouse MAb to EBV BZLF1 (Dako), and an MAb to EBV gp350 (C1, kindly provided by T. Sairenji).

Incubation:

Article Title: Toll-like Receptor 10 in Helicobacter pylori Infection
Article Snippet: .. Briefly, after antigen retrieval and inactivation of endogenous peroxidase activity, tissue sections were incubated with anti-TLR10 (H-165; Santa Cruz Biotechnology, Dallas, Texas) or rabbit polyclonal anti- H. pylori (DAKO, Copenhagen, Denmark) antibodies with diluting solution (DAKO) overnight at 4°C. .. Human NCI-N87 gastric cells were purchased from American Type Culture Collection (Manassas, Virginia).

Activity Assay:

Article Title: Toll-like Receptor 10 in Helicobacter pylori Infection
Article Snippet: .. Briefly, after antigen retrieval and inactivation of endogenous peroxidase activity, tissue sections were incubated with anti-TLR10 (H-165; Santa Cruz Biotechnology, Dallas, Texas) or rabbit polyclonal anti- H. pylori (DAKO, Copenhagen, Denmark) antibodies with diluting solution (DAKO) overnight at 4°C. .. Human NCI-N87 gastric cells were purchased from American Type Culture Collection (Manassas, Virginia).

Western Blot:

Article Title: The Soluble Recombinant Neisseria meningitidis Adhesin NadAΔ351–405 Stimulates Human Monocytes by Binding to Extracellular Hsp90
Article Snippet: .. No E. coli Ags were detected by Western immunoblot analysis with a rabbit polyclonal Ab risen against whole E. coli cells (DakoCytomation). ..

Marker:

Article Title: Involvement of the Tyro3 receptor and its intracellular partner Fyn signaling in Schwann cell myelination
Article Snippet: .. AntibodiesThe following antibodies were purchased: rabbit monoclonal anti-Tyro3 (5585S), rabbit monoclonal anti–neuronal marker GAP43 (8945S), rabbit monoclonal anti–68-kDa neurofilament NF68 (2835S), rabbit polyclonal anti-(pY527)Src (corresponding to pY531 in Fyn), and rabbit monoclonal anti–apoptotic marker active, cleaved caspase-3 (9579S) from Cell Signaling Technology (Danvers, MA); mouse monoclonal neuronal axon marker anti–βIII tubulin (MAB1195) from R & D Systems (Minneapolis, MN); mouse monoclonal anti-Fyn (05-436) and mouse monoclonal anti-(pS473)Akt (05-1003) from Merck-Millipore (Billerica, MA); rabbit polyclonal anti-Fyn (HPA023887) from Atlas Antibodies (Stockholm, Sweden); rabbit polyclonal anti–proliferating cell nuclear marker Ki67 (A0047) from Agilent Technology (Santa Clara, CA); mouse monoclonal anti-(pY416)Src (corresponding to pY420 in Fyn; sc-81521), mouse monoclonal anti-Akt1 (sc-5298), rabbit polyclonal anti-Oct6 (also called Scip; sc-133865), and rabbit polyclonal anti-Krox20 (also called Egr2; sc-20690) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit polyclonal anti-Tyro3 (ab109231) and mouse monoclonal anti-S100β (ab52642) from Abcam (Cambridge, United Kingdom); mouse monoclonal anti-MBP (SMI-94R-100) and monoclonal anti-GFAP (SMI-22R-100) from Covance (Princeton, NJ); anti-MPZ (also called P0) from MBL (Nagoya, Japan; rabbit polyclonal PD046) and Abnova (Taipei, Taiwan; goat polyclonal, PAB7332); mouse monoclonal anti–pan-sodium channel (K58/35) from Sigma-Aldrich (St. Louis, MO); mouse monoclonal anti-Rac1 (610652), mouse monoclonal anti-Cdc42 (610928), and mouse monoclonal anti–β-actin (612656) from Becton Dickinson (Franklin Lakes, NJ); mouse monoclonal anti–V5-epitope-tag (04434-94) from Nacalai Tesque (Kyoto, Japan); peroxidase-conjugated secondary antibodies from GE Healthcare (Fairfield, CT) and Nacalai Tesque; and fluorescence-labeled secondary antibodies from Life Technologies (Carlsbad, CA) and Wako Chemicals (Osaka, Japan). ..

Staining:

Article Title: Trypanosoma vivax Infections: Pushing Ahead with Mouse Models for the Study of Nagana. II. Immunobiological Dysfunctions
Article Snippet: .. Primary Ab against B220/CD45R, F4/80 and Ly49G2 were visualized using a Histofine Simple Stain MAX-PO kit (Histofine Biosciences inc, Cambridge, UK) and primary Ab against CD3 were visualized using peroxidase-labeled polymer for rabbit polyclonal Ab (EnVision, Dako, Carpinteria, CA, USA), according to the manufacturer's protocol. .. Color was developed with 3-Amino-9-EthylCarbazole (AEC chromogen; BD Pharmingen).

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    Agilent technologies antibody against ubiquitin
    Increased <t>ubiquitin-,</t> 20S proteasome- and CHOP-positive immunohistochemical staining in the rat retina exposed to rotenone and hydrogen peroxide. Vehicle ( a , 50% DMSO in distilled water), rotenone ( b , 2 nmol/eye), or hydrogen peroxide ( c , 1 μmol/eye, H 2 O 2 ) was intravitreally injected, and eyes were enucleated 1 and 24 hrs following injection. Each cross-sectioned retina was subjected to immunohistochemical staining for antibodies against ubiquitin (upper panel), 20S proteasome subunit (middle panel) and CHOP (lower panel). Each photograph shows the representative images of cross-sectioned retina following injection. GCL: the ganglion cell layer; IPL: the inner plexiform layer; INL: the inner nuclear layer. Arrows indicate the representative cells with positive immunostaining for each antibody. Each scale bar shows 20 μm. Note that this experiment was performed as part of our previous study published elsewhere (Kageyama et al ., PLoS One, 2019) 19 . The same control images are used in this figure.
    Antibody Against Ubiquitin, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies anti human von willebrand factor
    Distinctive features of the vascular and immune stroma of the four models of experimental malignant mesotheliomas Immunohistochemical evaluation of blood vessels density and size (vWf: von <t>Willebrand</t> factor), and of the macrophagic (ED1) and T-lymphocytic (CD3 and CD8) infiltration of the tumor stroma. The four models displayed distinctive characteristics: the M5-T2 tumors display a moderate blood vessel density (arrowheads) and a low T lymphocytic and macrophagic infiltration of the stroma (very low number of immunopositive cells on the ED1, CD3 and CD8 pictures); the F4-T2 and F5-T1 neoplasms are characterized by a moderate angiogenic activity (arrowheads) and a marked to severe infiltration of the tumor stroma by macrophages and particularly CD3+ T-lymphocytes, including CD8+ T-cells (asterisks: high number of immunopositive cells on the ED1, CD3 and CD8 pictures); the M5-T1 tumors are characterized by a high vascular density (arrowheads) and a low T lymphocytes and macrophages infiltration, particularly of the center of the nodules, with immune cells mostly located at the periphery of the neoplastic tissue (low number of immunopositive cells on the ED1, CD3 and CD8 pictures).
    Anti Human Von Willebrand Factor, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Agilent technologies antibodies against cd3
    ( A – C ) Immunofluorescence analyses labeling <t>CD3</t> and CD68 were performed on spinal cord sections upon completion of the experiment. ( A ) The number of T cells (CD3-immunopositive) was significantly lower in PBN animals than in mice of solvent and EAE groups. * p
    Antibodies Against Cd3, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Agilent technologies biotinylated pab rabbit anti human c3d
    (A) An illustration of the newly developed assay to measure non-proteolytically activated C3. (B) Schematic linear representation of native C3 [α-chain alone with indicated cleavage sites for convertases and Factor I appointed 1–3 (top of the figure) and the intact C3 molecule with a mark for the position of the thioester (below)], non-proteolytically activated C3 i.e., C3(x) (left) and C3 activated by convertases i.e., C3b (right) and their degradation fragments. The figure is adapted from Ekdahl et al. ( 25 ) with permission from the publisher. (C) Correlation between measured levels of C3(x) using <t>anti-C3d</t> or anti-C3c polyclonal antibodies for detection. (D) Measured C3(x) formation in plasma over time from 0 to 180 min.
    Biotinylated Pab Rabbit Anti Human C3d, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Increased ubiquitin-, 20S proteasome- and CHOP-positive immunohistochemical staining in the rat retina exposed to rotenone and hydrogen peroxide. Vehicle ( a , 50% DMSO in distilled water), rotenone ( b , 2 nmol/eye), or hydrogen peroxide ( c , 1 μmol/eye, H 2 O 2 ) was intravitreally injected, and eyes were enucleated 1 and 24 hrs following injection. Each cross-sectioned retina was subjected to immunohistochemical staining for antibodies against ubiquitin (upper panel), 20S proteasome subunit (middle panel) and CHOP (lower panel). Each photograph shows the representative images of cross-sectioned retina following injection. GCL: the ganglion cell layer; IPL: the inner plexiform layer; INL: the inner nuclear layer. Arrows indicate the representative cells with positive immunostaining for each antibody. Each scale bar shows 20 μm. Note that this experiment was performed as part of our previous study published elsewhere (Kageyama et al ., PLoS One, 2019) 19 . The same control images are used in this figure.

    Journal: Scientific Reports

    Article Title: Rotenone-induced inner retinal degeneration via presynaptic activation of voltage-dependent sodium and L-type calcium channels in rats

    doi: 10.1038/s41598-020-57638-y

    Figure Lengend Snippet: Increased ubiquitin-, 20S proteasome- and CHOP-positive immunohistochemical staining in the rat retina exposed to rotenone and hydrogen peroxide. Vehicle ( a , 50% DMSO in distilled water), rotenone ( b , 2 nmol/eye), or hydrogen peroxide ( c , 1 μmol/eye, H 2 O 2 ) was intravitreally injected, and eyes were enucleated 1 and 24 hrs following injection. Each cross-sectioned retina was subjected to immunohistochemical staining for antibodies against ubiquitin (upper panel), 20S proteasome subunit (middle panel) and CHOP (lower panel). Each photograph shows the representative images of cross-sectioned retina following injection. GCL: the ganglion cell layer; IPL: the inner plexiform layer; INL: the inner nuclear layer. Arrows indicate the representative cells with positive immunostaining for each antibody. Each scale bar shows 20 μm. Note that this experiment was performed as part of our previous study published elsewhere (Kageyama et al ., PLoS One, 2019) 19 . The same control images are used in this figure.

    Article Snippet: Serial cross sections (4-μm thickness) of the retina through the optic disk were prepared and incubated with a primary antibody against ubiquitin (1:200, rabbit polyclonal, Cat No. Z0458, Agilent Dako, Santa Clara, CA), 20S proteasome core subunits (1:1000, rabbit polyclonal, Cat No. PW8155, Enzo Life Sciences, Farmingdale, NY, USA) or CHOP (1:50, rabbit polyclonal, Cat No. sc-575, Santa Cruz, Santa Cruz, CA).

    Techniques: Immunohistochemistry, Staining, Injection, Immunostaining

    Distinctive features of the vascular and immune stroma of the four models of experimental malignant mesotheliomas Immunohistochemical evaluation of blood vessels density and size (vWf: von Willebrand factor), and of the macrophagic (ED1) and T-lymphocytic (CD3 and CD8) infiltration of the tumor stroma. The four models displayed distinctive characteristics: the M5-T2 tumors display a moderate blood vessel density (arrowheads) and a low T lymphocytic and macrophagic infiltration of the stroma (very low number of immunopositive cells on the ED1, CD3 and CD8 pictures); the F4-T2 and F5-T1 neoplasms are characterized by a moderate angiogenic activity (arrowheads) and a marked to severe infiltration of the tumor stroma by macrophages and particularly CD3+ T-lymphocytes, including CD8+ T-cells (asterisks: high number of immunopositive cells on the ED1, CD3 and CD8 pictures); the M5-T1 tumors are characterized by a high vascular density (arrowheads) and a low T lymphocytes and macrophages infiltration, particularly of the center of the nodules, with immune cells mostly located at the periphery of the neoplastic tissue (low number of immunopositive cells on the ED1, CD3 and CD8 pictures).

    Journal: Oncotarget

    Article Title: Characterization of increasing stages of invasiveness identifies stromal/cancer cell crosstalk in rat models of mesothelioma

    doi: 10.18632/oncotarget.24632

    Figure Lengend Snippet: Distinctive features of the vascular and immune stroma of the four models of experimental malignant mesotheliomas Immunohistochemical evaluation of blood vessels density and size (vWf: von Willebrand factor), and of the macrophagic (ED1) and T-lymphocytic (CD3 and CD8) infiltration of the tumor stroma. The four models displayed distinctive characteristics: the M5-T2 tumors display a moderate blood vessel density (arrowheads) and a low T lymphocytic and macrophagic infiltration of the stroma (very low number of immunopositive cells on the ED1, CD3 and CD8 pictures); the F4-T2 and F5-T1 neoplasms are characterized by a moderate angiogenic activity (arrowheads) and a marked to severe infiltration of the tumor stroma by macrophages and particularly CD3+ T-lymphocytes, including CD8+ T-cells (asterisks: high number of immunopositive cells on the ED1, CD3 and CD8 pictures); the M5-T1 tumors are characterized by a high vascular density (arrowheads) and a low T lymphocytes and macrophages infiltration, particularly of the center of the nodules, with immune cells mostly located at the periphery of the neoplastic tissue (low number of immunopositive cells on the ED1, CD3 and CD8 pictures).

    Article Snippet: Antibodies used for immunohistochemical analyses were: anti-human Ki67 (rabbit monoclonal SP6 clone 1/100, Spring Bioscience, Pleasanton, CA, USA) and anti-human von Willebrand Factor (rabbit polyclonal 1/400, Agilent Technologies, Les Ulis, France) with the IView Universal DAB detection kit (Roche Diagnostics) for revelation; and anti-rat ED1 antibody (MAB1435 1/100, EMD Millipore Corporation, Billerica, MA, USA) used as a pan-macrophage marker, mouse anti-CD3 (SM253P, Acris Antibodies, San Diego, USA), anti-CD8 (LS-B3665, LSBio France, 92000 Nanterre), with an anti-mouse secondary antibody and N-Histofine Simple Stain Mouse MAX Peroxidase (Nichirei Biosciences, Tokyo, Japan) as the detection reagent.

    Techniques: Immunohistochemistry, Activity Assay

    ( A – C ) Immunofluorescence analyses labeling CD3 and CD68 were performed on spinal cord sections upon completion of the experiment. ( A ) The number of T cells (CD3-immunopositive) was significantly lower in PBN animals than in mice of solvent and EAE groups. * p

    Journal: Scientific Reports

    Article Title: Probenecid arrests the progression of pronounced clinical symptoms in a mouse model of multiple sclerosis

    doi: 10.1038/s41598-017-17517-5

    Figure Lengend Snippet: ( A – C ) Immunofluorescence analyses labeling CD3 and CD68 were performed on spinal cord sections upon completion of the experiment. ( A ) The number of T cells (CD3-immunopositive) was significantly lower in PBN animals than in mice of solvent and EAE groups. * p

    Article Snippet: Four cross-sections from the spinal cord of each mouse (EAE n = 5, PBN n = 8, solvent n = 9, non-EAE n = 3) were immunolabelled with antibodies against CD3 (1:100; rabbit polyclonal antibody A0452; Agilent Technologies, Hamburg, Germany) and CD68 (1:100; rat monoclonal antibody ab53444; Abcam, Cambridge, UK).

    Techniques: Immunofluorescence, Labeling, Mouse Assay

    (A) An illustration of the newly developed assay to measure non-proteolytically activated C3. (B) Schematic linear representation of native C3 [α-chain alone with indicated cleavage sites for convertases and Factor I appointed 1–3 (top of the figure) and the intact C3 molecule with a mark for the position of the thioester (below)], non-proteolytically activated C3 i.e., C3(x) (left) and C3 activated by convertases i.e., C3b (right) and their degradation fragments. The figure is adapted from Ekdahl et al. ( 25 ) with permission from the publisher. (C) Correlation between measured levels of C3(x) using anti-C3d or anti-C3c polyclonal antibodies for detection. (D) Measured C3(x) formation in plasma over time from 0 to 180 min.

    Journal: Frontiers in Immunology

    Article Title: Assessment of the Role of C3(H2O) in the Alternative Pathway

    doi: 10.3389/fimmu.2020.00530

    Figure Lengend Snippet: (A) An illustration of the newly developed assay to measure non-proteolytically activated C3. (B) Schematic linear representation of native C3 [α-chain alone with indicated cleavage sites for convertases and Factor I appointed 1–3 (top of the figure) and the intact C3 molecule with a mark for the position of the thioester (below)], non-proteolytically activated C3 i.e., C3(x) (left) and C3 activated by convertases i.e., C3b (right) and their degradation fragments. The figure is adapted from Ekdahl et al. ( 25 ) with permission from the publisher. (C) Correlation between measured levels of C3(x) using anti-C3d or anti-C3c polyclonal antibodies for detection. (D) Measured C3(x) formation in plasma over time from 0 to 180 min.

    Article Snippet: The generated C3(x) was detected in three different ways: (1) by ELISA using monoclonal antibody (mAb) 4SD17.3 toward a neo-epitope in C3a for capture and biotinylated polyclonal (pAb) anti-C3d (Dako, Glostrup, Denmark) together with streptavidin-HRP (GE Healthcare, Uppsala, Sweden) for detection; (2) by Magpix® Luminex technology, using MagPlexC® Microspheres (Luminex, Bio-Rad Laboratories, Hercules, CA, USA) pre-coupled with mAb mouse anti-human C3a (4SD17.3) for capture (3 μg/1.25 × 106 beads) and 4 μg/mL biotinylated pAb anti-C3c (Dako, Glostrup, Denmark) followed by PE-conjugated streptavidin (Bio-Rad Laboratories, USA) for detection, and (3) using the same assay but biotinylated pAb rabbit anti-human C3d (Agilent Technologies, Inc., Santa Clara, CA, USA) instead of anti-C3c.

    Techniques: