Western Blot:Article Title: Kindlin-2 controls TGF-β signalling and Sox9 expression to regulate chondrogenesis
Article Snippet: The membrane was blocked in 5% non-fat milk in Tris-buffered saline/Tween 20 buffer; probed with primary antibodies, followed by incubation with secondary antibodies conjugated with horseradish peroxidase; and visualized using a Western Blotting Detection Kit (GE Healthcare, cat#: RPN2106). .. Antibodies used for western blotting in this study were Kindlin-1 (Sigma, SAB4200465, 1:1,000), Kindlin-3 (Cell Signaling, Cat# 13,843, 1:300), TGFβ R1 (Santa Cruz, sc-9048, 1:500), TGFβ R2 (Santa Cruz, sc-400, 1:500), pFAK (Santa Cruz, sc-11765, 1:300), Sox9 (Santa Cruz Biotechnology, sc-20095, 1:500), pSmad2/3 (Cell Signaling Technology, cat#: 8828S, 1:500), total Smad2/3 (Cell Signaling Technology, cat#: 8685S, 1:500), integrin β1 (BD, cat# 610468, 1:3,000), ILK (Cell Signaling, cat# I0783, 1:1,000), p38 MAPK (Cell Signaling, cat# 9,212, 1:1,000), phospho-p38 MAPK (Cell Signaling, cat# 9,216, 1:1,000), (Sigma-Aldrich, cat#: T-7451, 1:1,000), tubulin (Sigma-Aldrich, cat#: T-7451, 1:5,000), β-actin (Sigma-Aldrich, cat#: A5316, 1:4,000). ..
Article Title: Focal Adhesion Kinase Inhibition Ameliorates Burn Injury-Induced Chronic Pain in Rats.
Article Snippet: Burn injury-induced pain (BIP) is a significant global health concern, affecting diverse populations including children, military veterans, and accident victims.. Current pharmacotherapeutics for the management of BIP are associated with severe side effects including drug addiction, respiratory depression, sedation, and constipation posing significant barrier to their clinical utility.. In the present study, we have investigated the potential role of focal adhesion kinase (p-FAK) for the very first time in BIP and elucidated the associated underlying mechanisms.
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Membrane:Article Title: In-situ forming injectable GFOGER-conjugated BMSCs-laden hydrogels for osteochondral regeneration
Article Snippet: .. The membrane was incubated with primary antibodies against FAK (sc-558, dilution ratio 1:200, Santa Cruz Biotechnology, TX, USA), pFAK (sc-16662, dilution ratio 1:200, Santa Cruz Biotechnology), ERK (ab9363, dilution ratio 1:250, Abcam, Cambridge, UK), pERK (919301, dilution ratio 1:1000, Bio Legend, CA, USA), p38 (ab31828, dilution ratio 1:1000, Abcam), pp38 (ab4822, dilution ratio 1:1000, Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; MA5-15738, dilution ratio 1:1000, Thermo Fisher Scientific) at 4 °C overnight. .. The membranes were washed and incubated with secondary antibody (Anti-mouse IgG-HRP; 7076, dilution ratio 1:1000, Cell signaling) (Goat anti-rabbit IgG-HRP; sc-2004, dilution ratio 1:5000, Santa Cruz Biotechnology) (Goat anti-mouse IgG-HRP; sc-2005, dilution ratio 1:5000, Santa Cruz Biotechnology) (Rabbit anti-goat lgG H&L-HRP; ab6741, dilution ratio 1:5000, Abcam) conjugated to horseradish peroxidase (HRP) for 1 h at RT.
Article Title: In-situ forming injectable GFOGER-conjugated BMSCs-laden hydrogels for osteochondral regeneration.
Article Snippet: .. The membrane was incubated with primary antibodies against FAK (sc-558, dilution ratio 1:200, Santa Cruz Biotechnology, TX, USA), pFAK (sc-16662, dilution ratio 1:200, Santa Cruz Biotechnology), ERK (ab9363, dilution ratio 1:250, Abcam, Cambridge, UK), pERK (919301, dilution ratio 1:1000, Bio Legend, CA, USA), p38 (ab31828, dilution ratio 1:1000, Abcam), pp38 (ab4822, dilution ratio 1:1000, Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; MA5-15738, dilution ratio 1:1000, Thermo Fisher Scientific) at 4 °C overnight. .. The membranes were washed and incubated with secondary antibody (Anti-mouse IgG-HRP; 7076, dilution ratio 1:1000, Cell signaling) (Goat anti-rabbit IgG-HRP; sc-2004, dilution ratio 1:5000, Santa Cruz Biotechnology) (Goat anti-mouse IgG-HRP; sc-2005, dilution ratio 1:5000, Santa Cruz Biotechnology) (Rabbit anti-goat lgG H&L-HRP; ab6741, dilution ratio 1:5000, Abcam) conjugated to horseradish peroxidase (HRP) for 1 h at RT.
Incubation:Article Title: In-situ forming injectable GFOGER-conjugated BMSCs-laden hydrogels for osteochondral regeneration
Article Snippet: .. The membrane was incubated with primary antibodies against FAK (sc-558, dilution ratio 1:200, Santa Cruz Biotechnology, TX, USA), pFAK (sc-16662, dilution ratio 1:200, Santa Cruz Biotechnology), ERK (ab9363, dilution ratio 1:250, Abcam, Cambridge, UK), pERK (919301, dilution ratio 1:1000, Bio Legend, CA, USA), p38 (ab31828, dilution ratio 1:1000, Abcam), pp38 (ab4822, dilution ratio 1:1000, Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; MA5-15738, dilution ratio 1:1000, Thermo Fisher Scientific) at 4 °C overnight. .. The membranes were washed and incubated with secondary antibody (Anti-mouse IgG-HRP; 7076, dilution ratio 1:1000, Cell signaling) (Goat anti-rabbit IgG-HRP; sc-2004, dilution ratio 1:5000, Santa Cruz Biotechnology) (Goat anti-mouse IgG-HRP; sc-2005, dilution ratio 1:5000, Santa Cruz Biotechnology) (Rabbit anti-goat lgG H&L-HRP; ab6741, dilution ratio 1:5000, Abcam) conjugated to horseradish peroxidase (HRP) for 1 h at RT.
Article Title: In-situ forming injectable GFOGER-conjugated BMSCs-laden hydrogels for osteochondral regeneration.
Article Snippet: .. The membrane was incubated with primary antibodies against FAK (sc-558, dilution ratio 1:200, Santa Cruz Biotechnology, TX, USA), pFAK (sc-16662, dilution ratio 1:200, Santa Cruz Biotechnology), ERK (ab9363, dilution ratio 1:250, Abcam, Cambridge, UK), pERK (919301, dilution ratio 1:1000, Bio Legend, CA, USA), p38 (ab31828, dilution ratio 1:1000, Abcam), pp38 (ab4822, dilution ratio 1:1000, Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; MA5-15738, dilution ratio 1:1000, Thermo Fisher Scientific) at 4 °C overnight. .. The membranes were washed and incubated with secondary antibody (Anti-mouse IgG-HRP; 7076, dilution ratio 1:1000, Cell signaling) (Goat anti-rabbit IgG-HRP; sc-2004, dilution ratio 1:5000, Santa Cruz Biotechnology) (Goat anti-mouse IgG-HRP; sc-2005, dilution ratio 1:5000, Santa Cruz Biotechnology) (Rabbit anti-goat lgG H&L-HRP; ab6741, dilution ratio 1:5000, Abcam) conjugated to horseradish peroxidase (HRP) for 1 h at RT.
Chromatin Immunoprecipitation:Article Title:
Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used FAK (1:200; Santa Cruz Biotechnology; sc-558) pFAK (1:200; Santa Cruz Biotechnology; sc-16662) ERK (1:250; Abcam; ab9363) pERK (1:1000; Bio Legend; 919301) 3 nature portfolio | reporting sum m ary M arch 2021 p38 (1:1000; Abcam; ab31828) pp38 (1:1000; Abcam; ab4822) GAPDH (1:1000; Thermo Fisher Scientific; MA5-15738) Anti-mouse IgG-HRP (1:1000; Cell signaling; 7076) Goat anti-rabbit IgG-HRP (1:5000; Santa Cruz Biotechnology; sc-2004) Goat anti-mouse IgG-HRP(1:5000; Santa Cruz Biotechnology; sc-2005) Rabbit anti-goat lgG H&L-HRP(1:5000; Abcam; ab6741) Anti-Integrin beta 1 (1:100; Abcam; ab78502) Integrin α2 (1:100; Santa Cruz Biotechnology; sc-53353) Integrin α11 (1:100; Santa Cruz Biotechnology; sc-390091) Goat anti-mouse IgG H&L-DyLight 488 (1:500; Abcam; ab96879) goat anti-mouse IgG H&L (1:500; Abcam; ab97147) GFP (1:200; Santa Cruz Biotechnology; sc-9996) Anti-collagen type II (1:200; Abcam; ab34712), Collagen I (1:100; Novus Biologicals; NBP2-46874) Goat anti-Rabbit lgG (H+L) DyLight 594 (1:1000; Invitrogen; 35561) Validation Data provided in the manuscript. .. Animals and other research organisms Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research, and Sex and Gender in Research Laboratory animals Sprague-Dawley (SD) male rats (9-10 weeks old) were used in experiment.
Flow Cytometry:Article Title:
Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used FAK (1:200; Santa Cruz Biotechnology; sc-558) pFAK (1:200; Santa Cruz Biotechnology; sc-16662) ERK (1:250; Abcam; ab9363) pERK (1:1000; Bio Legend; 919301) 3 nature portfolio | reporting sum m ary M arch 2021 p38 (1:1000; Abcam; ab31828) pp38 (1:1000; Abcam; ab4822) GAPDH (1:1000; Thermo Fisher Scientific; MA5-15738) Anti-mouse IgG-HRP (1:1000; Cell signaling; 7076) Goat anti-rabbit IgG-HRP (1:5000; Santa Cruz Biotechnology; sc-2004) Goat anti-mouse IgG-HRP(1:5000; Santa Cruz Biotechnology; sc-2005) Rabbit anti-goat lgG H&L-HRP(1:5000; Abcam; ab6741) Anti-Integrin beta 1 (1:100; Abcam; ab78502) Integrin α2 (1:100; Santa Cruz Biotechnology; sc-53353) Integrin α11 (1:100; Santa Cruz Biotechnology; sc-390091) Goat anti-mouse IgG H&L-DyLight 488 (1:500; Abcam; ab96879) goat anti-mouse IgG H&L (1:500; Abcam; ab97147) GFP (1:200; Santa Cruz Biotechnology; sc-9996) Anti-collagen type II (1:200; Abcam; ab34712), Collagen I (1:100; Novus Biologicals; NBP2-46874) Goat anti-Rabbit lgG (H+L) DyLight 594 (1:1000; Invitrogen; 35561) Validation Data provided in the manuscript. .. Animals and other research organisms Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research, and Sex and Gender in Research Laboratory animals Sprague-Dawley (SD) male rats (9-10 weeks old) were used in experiment.
Magnetic Resonance Imaging:Article Title:
Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used FAK (1:200; Santa Cruz Biotechnology; sc-558) pFAK (1:200; Santa Cruz Biotechnology; sc-16662) ERK (1:250; Abcam; ab9363) pERK (1:1000; Bio Legend; 919301) 3 nature portfolio | reporting sum m ary M arch 2021 p38 (1:1000; Abcam; ab31828) pp38 (1:1000; Abcam; ab4822) GAPDH (1:1000; Thermo Fisher Scientific; MA5-15738) Anti-mouse IgG-HRP (1:1000; Cell signaling; 7076) Goat anti-rabbit IgG-HRP (1:5000; Santa Cruz Biotechnology; sc-2004) Goat anti-mouse IgG-HRP(1:5000; Santa Cruz Biotechnology; sc-2005) Rabbit anti-goat lgG H&L-HRP(1:5000; Abcam; ab6741) Anti-Integrin beta 1 (1:100; Abcam; ab78502) Integrin α2 (1:100; Santa Cruz Biotechnology; sc-53353) Integrin α11 (1:100; Santa Cruz Biotechnology; sc-390091) Goat anti-mouse IgG H&L-DyLight 488 (1:500; Abcam; ab96879) goat anti-mouse IgG H&L (1:500; Abcam; ab97147) GFP (1:200; Santa Cruz Biotechnology; sc-9996) Anti-collagen type II (1:200; Abcam; ab34712), Collagen I (1:100; Novus Biologicals; NBP2-46874) Goat anti-Rabbit lgG (H+L) DyLight 594 (1:1000; Invitrogen; 35561) Validation Data provided in the manuscript. .. Animals and other research organisms Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research, and Sex and Gender in Research Laboratory animals Sprague-Dawley (SD) male rats (9-10 weeks old) were used in experiment.
Biomarker Discovery:Article Title:
Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used FAK (1:200; Santa Cruz Biotechnology; sc-558) pFAK (1:200; Santa Cruz Biotechnology; sc-16662) ERK (1:250; Abcam; ab9363) pERK (1:1000; Bio Legend; 919301) 3 nature portfolio | reporting sum m ary M arch 2021 p38 (1:1000; Abcam; ab31828) pp38 (1:1000; Abcam; ab4822) GAPDH (1:1000; Thermo Fisher Scientific; MA5-15738) Anti-mouse IgG-HRP (1:1000; Cell signaling; 7076) Goat anti-rabbit IgG-HRP (1:5000; Santa Cruz Biotechnology; sc-2004) Goat anti-mouse IgG-HRP(1:5000; Santa Cruz Biotechnology; sc-2005) Rabbit anti-goat lgG H&L-HRP(1:5000; Abcam; ab6741) Anti-Integrin beta 1 (1:100; Abcam; ab78502) Integrin α2 (1:100; Santa Cruz Biotechnology; sc-53353) Integrin α11 (1:100; Santa Cruz Biotechnology; sc-390091) Goat anti-mouse IgG H&L-DyLight 488 (1:500; Abcam; ab96879) goat anti-mouse IgG H&L (1:500; Abcam; ab97147) GFP (1:200; Santa Cruz Biotechnology; sc-9996) Anti-collagen type II (1:200; Abcam; ab34712), Collagen I (1:100; Novus Biologicals; NBP2-46874) Goat anti-Rabbit lgG (H+L) DyLight 594 (1:1000; Invitrogen; 35561) Validation Data provided in the manuscript. .. Animals and other research organisms Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research, and Sex and Gender in Research Laboratory animals Sprague-Dawley (SD) male rats (9-10 weeks old) were used in experiment.
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