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iκb kinase alpha (ikkα) antibody  (Cell Signaling Technology Inc)
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Iκb Kinase Alpha (Ikkα) Antibody, supplied by Cell Signaling Technology 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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anti iκb kinase α  (Cell Signaling Technology Inc)
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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
Anti Iκb Kinase α, supplied by Cell Signaling Technology Inc, 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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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
Iκb Kinase β, supplied by Proteintech, 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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anti iκb kinase β ikkβ  (Cell Signaling Technology Inc)
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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
Anti Iκb Kinase β Ikkβ, 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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iκb kinase β  (Cell Signaling Technology Inc)
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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
Iκb Kinase β, 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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homozygous inhibitors of nuclear factor-κb (iκb) kinase (ikk)α-floxed mice m199006 c57bl/6 background  (Jackson Laboratory)
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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
Homozygous Inhibitors Of Nuclear Factor κb (Iκb) Kinase (Ikk)α Floxed Mice M199006 C57bl/6 Background, 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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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
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Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB <t>kinase</t> <t>α</t> (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.
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Image Search Results


Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB kinase α (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.

Journal: Journal of the American Heart Association

Article Title: Macrophage Lyn Kinase Is a Sex‐Specific Regulator of Post–Subarachnoid Hemorrhage Neuroinflammation

doi: 10.1161/jaha.124.039409

Figure Lengend Snippet: Figure 1. TLR4–Lyn interaction in hemorrhagic stroke. A, In vivo immunofluorescence of TLR4 (1: 500; red) and Lyn (1: 300; green) colocalization with DAPI in wild-type sham or SAH mouse brains on post-operative day 7, all scale bars=50 μm. B, In vitro and in vivo immunoprecipitation of TLR4 with coimmunoprecipitation of Lyn and MyD88 in microglia or mouse brain lysate; microglia incubated with red blood cells or PBS for 3 hours (in vitro) and sham or SAH on postoperative day 3 (in vivo) with 3 μg of mouse TLR4 antibody or 3 μg of mouse preimmune serum and 0.25 mg protein A/G beads, followed by immunoblot with TLR4 (1:1000), Lyn (1:250), and MyD88 (1:250) (n=3 for each group, representative blots shown). C, In vitro microglia and (BMM and in vivo mouse brain lysate protein expression of TLR4) (1:1000), Lyn (1:500), phospho-Lyn (1:500), MyD88 (1: 250), IκB kinase α (1: 250), nuclear factor-κB (1:250) and vinculin (1:1000) were measured. D and E, Flow cytometric analysis of macrophage TLR4 and Lyn expression in mice brains shown by t-distributed stochastic neighbor embedding and quantification of TLR4+Lyn+ macrophage cell numbers (n=4 males and n=4 females; ANOVA P<0.05, *P<0.05, **P<0.001 between groups). F, T- distributed stochastic neighbor embedding plots of sex-specific macrophage TLR4 and Lyn expression and quantification in microglia /BMM and female/male expression ratios (n=4 males and n=4 females; Student’s t test P<0.05, *P<0.05, **P<0.001 between groups). BMM indicates bone marrow–derived macrophages; IP, immunoprecipitation; MG, microglia; SAH, subarachnoid hemorrhage; and TLR4, Toll-like receptor 4.

Article Snippet: After blocking membrane was incubated with rabbit anti- Lyn (1:1000; CST, Catalog No. 2796S), rabbit anti- phosphoLyn (1:1000; Bioss, Catalog No. bs- 3257R), mouse antiTLR4 (1:1000; Invitrogen, Catalog No. 14–9041- 80), rabbit anti- GAPDH (1:1000; CST, Catalog No. 2118S), rabbit anti- Myd88 (1:1000; CST, Catalog No. 4283S), rabbit anti–nuclear factor- κB (1:1000; CST, Catalog No. 8242T) and rabbit anti–IκB kinase α (1:1000; CST, Catalog No. 61294) antibodies.

Techniques: In Vivo, Immunofluorescence, In Vitro, Immunoprecipitation, Incubation, Western Blot, Expressing, Derivative Assay