polyclonal goat anti mouse tlr2 ab  (R&D Systems)

Journal: European journal of immunology

Article Title: TLR2 on CD4+ and CD8+ T cells promotes control of Mycobacterium tuberculosis infection.

doi: 10.1002/eji.202350715

Figure Lengend Snippet: Figure 1. Deletion of exon 3 of Tlr2 in CD4+ and CD8+ T cells in Tlr2fl/flxCd4cre/cre mice. (A) Location of LoxP sites around exon 3 of Tlr2 in Tlr2fl/fl

Article Snippet: Blots were either incubated with polyclonal goat anti-mouse TLR2 Ab (R&D Systems; AF1530) or β-actin (R&D Systems; MAB8929), followed by HRP-conjugated donkey anti-goat IgG(H+L) mAb (Jackson) and bands detected by chemiluminescence using Pierce ECL Plus substrate (Fisher).

Techniques:

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A and B) Western blot shows TLR1 and TLR6 in NK cells. (C and D) Immunoprecipitations were performed with anti-TLR2 in non-stimulated (lane a) and LPG-stimulated NK cells after 1 h of co-incubation (lane b). (C) Immunoprecipitates were subjected to Western blotting and probed with anti-TLR1 and (D) anti-TLR6 antibodies, respectively. A representative immunoblot from four different experiments is shown. (E) NK cell lysates in non-stimulated and LPG-stimulated NK cells were immunoprecipitated with anti-TLR2, anti-MyD88, anti-IRAK-1 and anti-TRAF-6 antibodies and Western blotted with anti-MyD88, anti-IRAK-1, anti-TRAF-6 and anti-IKK-γ antibodies. (F) Nuclear translocation of p50 and p65 NF-κB isoforms were analyzed in non-stimulated and LPG-stimulated NK cells. (G) Phosphorilation of pIKK-α/β and pIκB-α were analyzed in non-stimulated NK cells (Non-stim) or in cells stimulated with PMA (15 min) or LPG (15, 30, 45 and 60 min). A representative immunoblot from two different experiments is show.

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Western Blot, Incubation, Immunoprecipitation, Translocation Assay

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) Representative flow CD56 + /CD3 - expression in membranes of NK cells; right image: NK cells were gated and analyzed for TLR2 (TLR2 + /CD56 + ) expression. (B) Representative histograms showing the MFI levels of TLR2 expression on NK cells from control and patients (LCL and DCL). (C) Quantitation of TLR2 expression on NK cells. □ Non-stimulated cells, ▪ LPG-stimulated cells. Data are representative of healthy controls (n = 21), LCL patients (n = 28) and DCL patients (n = 6). These results are the mean ±SEM. *p≤0.05 was considered significant.

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Expressing, Control, Quantitation Assay

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) Analysis according to disease form. (B) TLR2, TLR1 and TLR6 expression according to age (≤25 or ≥26 years) in LCL patients. □ Non-stimulated NK cells, ▪ LPG-stimulated NK cells. Cell surface expression is indicated by MFI. These results are the mean ±SEM. *Significant differences were observed between LCL and DCL patients for all TLRs in LPG-stimulated and non-stimulated cells.

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Expressing

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) Double immunohistochemistry staining of TLR expression (TLR1, TLR2 and TLR6) on NK cells (CD57). (B) Number of NK cells expressing TLRs per mm 2 . Results are the mean ±SEM. Scale bar = 50 µm. Black arrows show double positive cells.

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) Representative flow cytometry dot plot (TLR2 vs. CD56) and histogram of TLR2 expression in NK cells. (B) This gate was subsequently analyzed for TLR2 expression on NK-cell subsets. Dot plot (CD16 + /CD56 + ) and histogram for CD56 bright (blue box) and CD56 dim (green box) NK cells. (C) TLR1 expression. (D) TLR2 expression. (E) TLR6 expression. □ Non-stimulated NK cells; ▪ LPG-stimulated NK cells. Lane 1: CD56 dim ; lane 2: CD56 bright NK cells. Cell surface expression is indicated by MFI. Results are the mean ±SEM. *Significant differences were observed between NK cell subsets of LCL and DCL patients for all 3 TLRs in LPG-stimulated and non-stimulated cells. LCL patients (n = 9), DCL patients (n = 4) and controls (n = 4).

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Flow Cytometry, Expressing

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) IFN-γ production of peripheral blood NK cells [control subjects (n = 21), LCL patients (n =  28) and DCL patients (n = 6)]. Analysis in LCL patients [female (n = 11) and male (n = 17)] according to: (B) gender; (C) disease duration; (D) age. (E) Double immunohistochemical labelling (CD57 + /IFN-γ + ) in lesions of patients (LCL and DCL) showed redish-brown staining generated by the combination of a red AP substrate used for NK cells and DAB Black used for IFN-γ staining. □ Non-stimulated NK cells. ▪ LPG-stimulated NK cells. Mean ±SEM is shown. *p≤0.05 was considered significant. Scale bar  = 50 µm. Black arrows show double positive cells.

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Control, Immunohistochemical staining, Staining, Generated

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: (A) TNF-α production in peripheral blood [control subjects (n = 21), LCL patients (n =  28) and DCL patients (n = 6)]. (B) Analysis of TNF-α production of LCL patients according to gender [female (n = 11) and male (n = 17)]. (C) Double immunohistochemistry (CD57 + /TNF-α + ) staining of lesions of LCL and DCL patients showed dark green staining induced by the combination of a green substrate (Stay Green/AP) used for NK cells and DAB (brown) used for TNF-α staining. □ Non-stimulated NK cells, ▪ LPG-stimulated NK cells. Mean ±SEM is shown. *p≤0.05 was considered significant. Scale bar  = 50 µm. Double positive cells CD57 + /TNF-α + (black arrows) and single positive cells CD57 - /TNF-α + (red arrows).

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Control, Immunohistochemistry, Staining

Journal: PLoS ONE

Article Title: NK Cell Activity Differs between Patients with Localized and Diffuse Cutaneous Leishmaniasis Infected with Leishmania mexicana: A Comparative Study of TLRs and Cytokines

doi: 10.1371/journal.pone.0112410

Figure Lengend Snippet: Transcript expression by quantitative real time PCR in non-stimulated (NS) and LPG-stimulated NK cells (S).

Article Snippet: The antibodies used for this analysis included: PE-conjugated anti-CD56, PE-Cy TM 5-anti-CD16, PE-Cy TM 5 mouse (IgG1k) and PE mouse (IgG1k) for isotype controls, all from BD-Pharmingen; rabbit polyclonal IgG TLR6 (H-90), goat polyclonal IgG TLR1 (N-20), goat polyclonal IgG TLR2 (N-17) all from Santa Cruz Biotechnology; FITC rabbit anti-goat IgG (H+L) conjugated, FITC goat anti-rabbit IgG (H+L) from Zymed and FITC goat IgG isotype control from Coulter.

Techniques: Expressing, Real-time Polymerase Chain Reaction