normal goat igg biotinylated control Search Results


streptavidin  (R&D Systems)
94
R&D Systems streptavidin
Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and <t>Streptavidin-PE.</t> (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.
Streptavidin, supplied by R&D Systems, 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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normal goat igg  (R&D Systems)
92
R&D Systems normal goat igg
Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and <t>Streptavidin-PE.</t> (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.
Normal Goat Igg, supplied by R&D Systems, 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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biotinylated normal goat igg  (Genzyme)
90
Genzyme biotinylated normal goat igg
Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and <t>Streptavidin-PE.</t> (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.
Biotinylated Normal Goat Igg, supplied by Genzyme, 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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biotinylated goat igg for isotype controls  (Jackson Laboratory)
90
Jackson Laboratory biotinylated goat igg for isotype controls
Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and <t>Streptavidin-PE.</t> (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.
Biotinylated Goat Igg For Isotype Controls, supplied by Jackson Laboratory, 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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Image Search Results


Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and Streptavidin-PE. (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.

Journal: European journal of immunology

Article Title: Comparison of stable human Treg and Th clones by transcriptional profiling.

doi: 10.1002/eji.200838807

Figure Lengend Snippet: Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and Streptavidin-PE. (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.

Article Snippet: For surface LAP expression, cells activated for 24 h with anti-CD3 and anti-CD28 antibodies in the presence of IL-2 (160 IU/mL) were labeled with a biotinylated polyclonal anti-LAP antibody or an isotype control (R&D Systems, BAF246 and BAF108, respectively), followed by incubation with Streptavidin coupled to PE (BD Pharmingen).

Techniques: Clone Assay, Membrane, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Control, Western Blot, Activation Assay, Recombinant, Co-Culture Assay