il 6 Search Results


mouse interleukin 6 il 6 elisa kit  (Shanghai Korain Biotech Co Ltd)
94
Shanghai Korain Biotech Co Ltd mouse interleukin 6 il 6 elisa kit
<t>Interleukin-6</t> <t>(IL-6)</t> concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)
Mouse Interleukin 6 Il 6 Elisa Kit, supplied by Shanghai Korain Biotech Co Ltd, 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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gene exp il6 hs00174131 m1  (Thermo Fisher)
99
Thermo Fisher gene exp il6 hs00174131 m1
<t>Interleukin-6</t> <t>(IL-6)</t> concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)
Gene Exp Il6 Hs00174131 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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il 6  (Procell Inc)
86
Procell Inc il 6
<t>Interleukin-6</t> <t>(IL-6)</t> concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)
Il 6, supplied by Procell 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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immunohistochemical interleukin il  (Servicebio Inc)
86
Servicebio Inc immunohistochemical interleukin il
Histology and <t>IL-2</t> expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)
Immunohistochemical Interleukin Il, supplied by Servicebio 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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il 6  (R&D Systems)
93
R&D Systems il 6
Histology and <t>IL-2</t> expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)
Il 6, supplied by R&D Systems, 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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il 6  (Miltenyi Biotec)
93
Miltenyi Biotec il 6
Histology and <t>IL-2</t> expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)
Il 6, supplied by Miltenyi Biotec, 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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human recombi  (Miltenyi Biotec)
95
Miltenyi Biotec human recombi
Histology and <t>IL-2</t> expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)
Human Recombi, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human ku  (Danaher Inc)
99
Danaher Inc recombinant human ku
FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to <t>recombinant</t> RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).
Recombinant Human Ku, supplied by Danaher Inc, 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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il 6  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology il 6
FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to <t>recombinant</t> RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).
Il 6, supplied by Santa Cruz Biotechnology, 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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il 6 human elisa kit  (Danaher Inc)
94
Danaher Inc il 6 human elisa kit
FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to <t>recombinant</t> RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).
Il 6 Human Elisa Kit, supplied by Danaher Inc, 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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interleukin 6  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc interleukin 6
FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to <t>recombinant</t> RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).
Interleukin 6, 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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Image Search Results


Interleukin-6 (IL-6) concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)

Journal: ADMET & DMPK

Article Title: Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells

doi: 10.5599/admet.3149

Figure Lengend Snippet: Interleukin-6 (IL-6) concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)

Article Snippet: Other materials used in this study were Dulbecco’s modified eagle medium (DMEM) - high glucose, foetal bovine serum (FBS), Antibiotic-Antimycotic (ABAM), polyethylene glycol (PEG) 6000, trehalose (Sigma-Aldrich; Merck), phosphate-buffered saline (PBS), sodium periodate (NaIO 4 ) (Sigma-Aldrich), bovine serum albumin (BSA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT), 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), paraformaldehyde (PFA), kit BCA Assay (PierceTM BCA Protein Assay Kit, Thermo Scientific), PKH67 Green Fluorescent Cell Linker Kits (Sigma-Aldrich; Merck), and Mouse Interleukin-6 (IL-6) ELISA Kit (BT Lab, Cat. No. E0049Mo).

Techniques: Concentration Assay, Standard Deviation

Histology and IL-2 expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Histology and IL-2 expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Expressing, Control, Immunohistochemical staining, Staining

Immunohistochemical overview of anorectal cytokine expression. Expression of A IL-2, B IL-6, C , and D TNFα in anorectal tissue in each group detected by immunohistochemical staining (IHC, ×50; a model group, b control group)

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Immunohistochemical overview of anorectal cytokine expression. Expression of A IL-2, B IL-6, C , and D TNFα in anorectal tissue in each group detected by immunohistochemical staining (IHC, ×50; a model group, b control group)

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Immunohistochemical staining, Expressing, Staining, Control

Relative mRNA expression of pro-inflammatory cytokines A IL-2, B IL-6, and C TNFα in anorectal tissue in each group. * P < 0.05 vs control group

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Relative mRNA expression of pro-inflammatory cytokines A IL-2, B IL-6, and C TNFα in anorectal tissue in each group. * P < 0.05 vs control group

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Expressing, Control

Western blotting for detecting protein expression of IL-2, IL-6, and TNFα

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Western blotting for detecting protein expression of IL-2, IL-6, and TNFα

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Western Blot, Expressing

IL-2, IL-6, and TNFα protein levels in anorectal tissue are significantly elevated in the model group compared to the control group (* P < 0.05). A IL-2, B IL-6, and C TNFα protein expression in anorectal tissue. * P < 0.05 vs control group

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: IL-2, IL-6, and TNFα protein levels in anorectal tissue are significantly elevated in the model group compared to the control group (* P < 0.05). A IL-2, B IL-6, and C TNFα protein expression in anorectal tissue. * P < 0.05 vs control group

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Control, Expressing

FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to recombinant RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).

Journal: Journal of Biological Chemistry

Article Title: Loading of the Nonhomologous End Joining Factor, Ku, on Protein-occluded DNA Ends

doi: 10.1074/jbc.m611125200

Figure Lengend Snippet: FIGURE 7. The DNA binding activity of Ku at RAG bound signal ends. A, a signal end complex is defined here as RAG1, RAG2, and HMG1 bound to two- oligonucleotide DNA duplexes, one containing the 12-RS and the other con- taining the 23-RS. A streptavidin-biotin complex was used to block the ends of the DNA duplexes distal to the site of cleavage. B, approximately 10 ng/l of purified RAG1 and RAG2 and 425 nM HMG1 were incubated with 0.4 nM radiolabeled23-RS-containingand10nM12-RS-containingDNAfragmentsat 37 °C for 10 min. 5 M streptavidin was added at 25 °C for 5 min prior to addition of 25 nM Ku. Antibody supershifts required an additional 10-min room temperature incubation step with either 0.2 g of the monoclonal anti- body to Ku or 1 l of a polyclonal antisera recognizing the maltose binding domain fused to recombinant RAG proteins. The inferred composition of each of the generated species is indicated to the side of the panel: species I, 23-RS; species II, HMG1-bound 23-RS; species III, RAGs and HMG1 bound to the 23-RS; species IV, SEC; species V, streptavidin-blocked SEC (upper arrow indi- cates -MBP supershift); species VI, Ku bound to incompletely formed SEC (upper arrow indicates -Ku supershift).

Article Snippet: The relative amounts of Ku and histoneH3 in each of the excised complexes were determined by semi- quantitative Western analysis probing with a polyclonal rabbit antibody raised against native, recombinant human Ku and a polyclonal antibody against histone H3 (Ab1791; Abcam) using fluorescent detection and a Typhoon imager (GE Healthcare).

Techniques: Binding Assay, Activity Assay, Blocking Assay, Purification, Incubation, Recombinant, Generated