toll box Search Results


gene exp b2m mm00437762 m1  (Thermo Fisher)
99
Thermo Fisher gene exp b2m mm00437762 m1
Gene Exp B2m Mm00437762 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti hmgb1  (Proteintech)
96
Proteintech anti hmgb1
Anti Hmgb1, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti tlr2 monoclonal antibody mab  (OriGene)
99
OriGene mouse anti tlr2 monoclonal antibody mab
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Mouse Anti Tlr2 Monoclonal Antibody Mab, supplied by OriGene, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 1 article reviews
mouse anti tlr2 monoclonal antibody mab - by Bioz Stars, 2026-04
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co 80539 hach call toll free 1 800 525 5940  (Danaher Inc)
86
Danaher Inc co 80539 hach call toll free 1 800 525 5940
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Co 80539 Hach Call Toll Free 1 800 525 5940, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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tls matlab toll box  (MathWorks Inc)
90
MathWorks Inc tls matlab toll box
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Tls Matlab Toll Box, supplied by MathWorks Inc, 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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toll box  (MathWorks Inc)
90
MathWorks Inc toll box
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Toll Box, supplied by MathWorks Inc, 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/toll box/product/MathWorks Inc
Average 90 stars, based on 1 article reviews
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gene exp tbp hs00427620 m1  (Thermo Fisher)
99
Thermo Fisher gene exp tbp hs00427620 m1
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Gene Exp Tbp Hs00427620 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti foxc2 antibody  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti foxc2 antibody
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Anti Foxc2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pdonr223 atf4 wt tol2 uas atf4 2a mcherry spliced xbp1 xbp1s adgene plasmid 63680  (Addgene inc)
93
Addgene inc pdonr223 atf4 wt tol2 uas atf4 2a mcherry spliced xbp1 xbp1s adgene plasmid 63680
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Pdonr223 Atf4 Wt Tol2 Uas Atf4 2a Mcherry Spliced Xbp1 Xbp1s Adgene Plasmid 63680, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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abl kinase inhibitors imatinib  (Novartis)
90
Novartis abl kinase inhibitors imatinib
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Abl Kinase Inhibitors Imatinib, supplied by Novartis, 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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select tcs bait box for topical application of fipronil  (Tick Box Technology Corporation)
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Tick Box Technology Corporation select tcs bait box for topical application of fipronil
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Select Tcs Bait Box For Topical Application Of Fipronil, supplied by Tick Box Technology Corporation, 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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gene exp actb hs01060665 g1  (Thermo Fisher)
99
Thermo Fisher gene exp actb hs01060665 g1
(a) FACS analysis of CEACAM1 and <t>TLR2</t> (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.
Gene Exp Actb Hs01060665 G1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


(a) FACS analysis of CEACAM1 and TLR2 (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.

Journal: PLoS ONE

Article Title: Soluble CEACAM8 Interacts with CEACAM1 Inhibiting TLR2-Triggered Immune Responses

doi: 10.1371/journal.pone.0094106

Figure Lengend Snippet: (a) FACS analysis of CEACAM1 and TLR2 (black lines) expression on the cell surface of NHBE cells. Grey lines represent isotype-matched control mAb staining. (b, c and d) ELISA for IL8 and IL6, respectively, in supernatants of NHBE cells incubated with Pam 3 Cys alone, with Pam 3 Cys plus soluble CEACAM8-Fc and rat-CEACAM1-Fc, respectively (100 ng/ml each) or left untreated for 16 h. (e) 50 ng purified protein of CEACAM8-Fc and ratCEACAM1-Fc were analyzed by SDS-PAGE and stained by Coomassie blue to demonstrate equal amount and integrity of the used fusion proteins. (f) FACS analysis CEACAM1 and TLR2 expression (black lines) on the surface of A549 cells. Grey lines represent the isotype-matched control. (g) IL-8 released into the supernatants of A549 cells pre-incubated for 1 h with anti-CEACAM1 (mAb 18/20) or left untreated followed by stimulation with Pam 3 Cys alone or with Pam 3 Cys and CEACAM8-Fc and ratCEACAM1-Fc, respectively (100 ng/ml) for 16 h. Samples were measured by commercial ELISA. IL8 secretion following treatment of the cells with Pam3Cys served as positive control. (h) A549 cells were challenged either with CEACAM8-Fc or with mAb 18/20 (20 ng/ml). The IgG antibody was used as control. Cell lysates were assessed after CEACAM1 immunoprecipitation with anti-phospho tyrosine mAb 4G10 (upper panel). Then membrane was stripped and re-probed with anti-CEACAM1 mAb (lower panel) to measure the amount of precipitated CEACAM1. Data presented in (a, e, f and h) are from one experiment representative of three independent experiments, and the data presented in (b, c, d and g) are mean ± s.e.m. of three different experiments performed in triplicates, * P<0.05.

Article Snippet: After blocking with 10% FCS in PBS for 10 min, 0.5×10 6 cells were incubated with 10 µg/ml mouse anti-TLR2 monoclonal antibody (mAb) (Acris Antibodies GmbH, Herford, Germany) or 3.5 µg/ml anti-CEACAM1 mAb 18/20 (B.B.

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay, Incubation, Purification, SDS Page, Positive Control, Immunoprecipitation