primary antibodies trka  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Designed NGF mimetics with reduced nociceptive signatures in neurons

doi: 10.1101/2025.04.14.648806

Figure Lengend Snippet: Dose-response curves for pERK (middle) and pAKT (right) activation measured via phosphoflow for different backbone classes of all designs (colored) (A) Large designs (B) DHR04-based (C) 6-helical bundles (D) 6-helical short connector bundles (E) direct fusions and (F) direct long fusions. Error bars represent SD from three independent biological repeats. (G) Activation strength for the individual designs correlate for ERK and AKT phosphorylation. (H) Designs Nt-231 (top) and Nt-237 (bottom) were tested for specific TrkA pathway activation using a SRE-luciferase reporter gene and treatment with different concentrations of agonists. Designs did not lead to pathway activation of TrkB or TrkC. Error bars represent SEM from five independent biological repeats. (I) Time-course of TrkA receptor endocytosis for various concentrations of Nt-231 (top), Nt-237 (middle) agonists or NGF (bottom) using a BRET assay. Error bars represent SEM from five independent biological repeats.

Article Snippet: After blocking the membrane in 5% milk, 0.1% Tween, 10 mM Tris at pH 7.6, 100 mM NaCl, primary antibodies TrkA (Cell Signaling, Cat. 30697 at 1:1000), ERK (Cell Signaling, Cat. 9107), pERK (Cell Signaling, Cat. 4695), HSC70 (Santa Cruz, Cat. sc-7298; at 1:10.000), and Beta-Actin (Sigma, Cat. A1978 at 1:10,000) were added.

Techniques: Activation Assay, Phospho-proteomics, Luciferase, Bioluminescence Resonance Energy Transfer

Journal: Science translational medicine

Article Title: Lipid nanoparticles and siRNA targeting plasminogen provide lasting inhibition of fibrinolysis in mouse and dog models of hemophilia A

doi: 10.1126/scitranslmed.adh0027

Figure Lengend Snippet: (A) Mice were injected with a single dose of siLuc (gray) or siPlg (teal blue). Plg mRNA in liver tissue was measured using qPCR, normalized against the housekeeping gene, Ppia. (B) Representative Western blot against plasminogen, where each lane contains the plasma from an individual mouse in either treatment group. The triangular marker indicates the expected molecular weight of plasminogen (92 kDa). (C) Plasminogen protein in plasma measured in a portion of the mice enrolled at each time point using densitometry, normalized to a loading control, and graphed relative to untreated mice. (D) Plasminogen protein in plasma quantified after administration of siPlg to dogs at 0.027 (purple, n = 3), 0.054 (green, n = 2), or 0.54 (teal blue, n = 1) mg siRNA/kg body weight. (E) Representative Western blot against plasminogen, where each lane contains plasma collected from a single WT dog at a different time point before or after siPlg administration at 0.054 mg/kg. Data are presented as mean ± SEM and were analyzed by two-tailed unpaired Student’s t test (A) or by two-way ANOVA (C); *P < 0.05.

Article Snippet: The membranes were treated with a primary antibody against plasminogen (1:1000; SAPG-AP; confirmed cross-reactivity: human, rat, mouse, rabbit, canine, and pig; Affinity Biologicals), washed, and treated with horseradish peroxidase–labeled anti-host secondary antibody (1:15,000; Abcam).

Techniques: Injection, Western Blot, Clinical Proteomics, Marker, Molecular Weight, Control, Two Tailed Test

Journal: Science translational medicine

Article Title: Lipid nanoparticles and siRNA targeting plasminogen provide lasting inhibition of fibrinolysis in mouse and dog models of hemophilia A

doi: 10.1126/scitranslmed.adh0027

Figure Lengend Snippet: Samples were collected from two HA dogs at baseline (green), within 4 hours of TXA (10 mg/kg iv, orange), within 3 weeks of siPlg administration without TXA (teal), or after administration of both siPlg and TXA (purple). (A) Representative TEG curve tracings using plasma from two HA dogs (top and bottom graphs) with added tPA. (B) Clot lysis time within 180-min monitoring period, values represent mean, and error bars represent ± SEM. (C) Thrombin generation in plasma collected weekly from two HA dogs or pooled plasma from WT dogs (WT, gray). (D) Correlation between plasminogen protein in plasma and thrombin generation peak (open and closed markers distinguish the two siPlg-treated dogs). (E) Plasmin generation in plasma collected weekly from two HA dogs or pooled plasma from WT dogs. (F) Correlation between plasma plasminogen and plasmin generation peak.

Article Snippet: The membranes were treated with a primary antibody against plasminogen (1:1000; SAPG-AP; confirmed cross-reactivity: human, rat, mouse, rabbit, canine, and pig; Affinity Biologicals), washed, and treated with horseradish peroxidase–labeled anti-host secondary antibody (1:15,000; Abcam).

Techniques: Clinical Proteomics, Lysis

Journal: Science translational medicine

Article Title: Lipid nanoparticles and siRNA targeting plasminogen provide lasting inhibition of fibrinolysis in mouse and dog models of hemophilia A

doi: 10.1126/scitranslmed.adh0027

Figure Lengend Snippet: (A) Plasma plasminogen (left y axis) quantified during 9 months of repeat administration of siPlg. The weight of mice (right y axis) was tracked over this period during administration of siLuc (gray) and siPlg (orange). (B) Periodontal bone loss measured by the distance between the cementoenamel junction and the alveolar bone crest (orange lines) in samples stained with methylene blue from mice administered siLuc (n = 6) or siPlg (n = 7) for 9 months compared to untreated (Unt WT) age-matched WT controls (purple line). Scale bar, 2 mm. (C to F) Inflammatory markers IL-6 (C and D) and TNF-α (E and F) were measured using qPCR in liver tissue or enzyme-linked immunosorbent assay in plasma from siLuc-treated (n = 8) or siPlg-treated (n = 10) mice compared to untreated (Unt) controls (purple line). (G) Immunohistochemistry against fibrin(ogen) in microscopy images of liver tissue of siPlg-treated mice showed inflammatory infiltrates but no fibrinous lesions. Scale bar, 100 μm. (H) Immunohistochemistry against fibrin(ogen) in clots formed ex vivo in whole blood from mice treated with siLuc (left) or siPlg (right). Scale bar, 25 μm. For all graphs, values represent mean ± SEM, ns indicates difference not statistically significant (P > 0.05), analyzed by two-tailed unpaired Student’s t test.

Article Snippet: The membranes were treated with a primary antibody against plasminogen (1:1000; SAPG-AP; confirmed cross-reactivity: human, rat, mouse, rabbit, canine, and pig; Affinity Biologicals), washed, and treated with horseradish peroxidase–labeled anti-host secondary antibody (1:15,000; Abcam).

Techniques: Clinical Proteomics, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Microscopy, Ex Vivo, Two Tailed Test

Journal: Science translational medicine

Article Title: Lipid nanoparticles and siRNA targeting plasminogen provide lasting inhibition of fibrinolysis in mouse and dog models of hemophilia A

doi: 10.1126/scitranslmed.adh0027

Figure Lengend Snippet: (A) Schematic showing SVP and TVT injury models. (B) Plasminogen in HA mice at the time of TVT bleed. (C and D) Blood loss in microliters of blood per gram of body weight (C) and bleeding time (D) after TVT in WT, untreated HA, and treated HA mice (n = 5 to 7). (E and F) Blood loss in microliters of blood per gram of body weight (E) and bleeding time (F) to end of observation period at 40 min (gray dashed line) after SVP (n = 7 to 10). Values represent mean ± SEM; *P < 0.05, **P < 0.01, and ns indicates not significant (P > 0.05), analyzed by one-way ANOVA.

Article Snippet: The membranes were treated with a primary antibody against plasminogen (1:1000; SAPG-AP; confirmed cross-reactivity: human, rat, mouse, rabbit, canine, and pig; Affinity Biologicals), washed, and treated with horseradish peroxidase–labeled anti-host secondary antibody (1:15,000; Abcam).

Techniques:

Journal: Science translational medicine

Article Title: Lipid nanoparticles and siRNA targeting plasminogen provide lasting inhibition of fibrinolysis in mouse and dog models of hemophilia A

doi: 10.1126/scitranslmed.adh0027

Figure Lengend Snippet: HA dogs were administered five doses of siPlg over 15 weeks. Vertical dashed lines indicate days of an administration. (A) Plasminogen in plasma quantified using densitometry of Western blots. Values represent mean ± SEM. (B) Annualized bleeding rate extrapolated from 10 months before treatment and the 4-month period of siPlg treatment in two HA dogs (circle and diamond each indicate an individual dog).

Article Snippet: The membranes were treated with a primary antibody against plasminogen (1:1000; SAPG-AP; confirmed cross-reactivity: human, rat, mouse, rabbit, canine, and pig; Affinity Biologicals), washed, and treated with horseradish peroxidase–labeled anti-host secondary antibody (1:15,000; Abcam).

Techniques: Clinical Proteomics, Western Blot

Journal: Journal of Inflammation Research

Article Title: Augmented Cornus officinalis Levels in Liuwei Dihuang Decoction Inhibits Nucleus Pulposus Cell Pyroptosis to Enhance Therapeutic Efficacy Against Intervertebral Disc Degeneration

doi: 10.2147/JIR.S465690

Figure Lengend Snippet: LWDHD formulations activate the NRF2 antioxidant pathway in IVDs of IVDD mice. ( A and B ) IF staining and corresponding qualification of NRF2 (S40) in IVDs of IVDD mice after treatment with LWDHD formulation for 1 week. White arrows indicate high expression of NRF2 (S40) in NP tissues. ( C and D ) IHC staining and corresponding qualification of KEAP1 in IVDs of IVDD mice. Black arrows indicate high expression of KEAP1 in NP tissues. ( E – H ) IF staining and corresponding qualification of p-I-κB ( E and F ) and p-P65 ( G and H ) in IVDs of IVDD mice. Data are shown as mean ± SEM (n = 6). *Denotes a significant difference compared with the Sham group (** p < 0.01), # Means a significant difference compared with the Vehicle group ( # p < 0.05, ## p < 0.01), + Indicates a significant difference compared with the LWDHD group ( ++ p < 0.01), & Means a significant difference compared with the 5 × CO group ( && p < 0.01).

Article Snippet: Primary antibodies against Tyrosine kinase A (TrkA), and NRF2 (S40) were obtained from HuaBio (Hangzhou, China).

Techniques: Staining, Formulation, Expressing, Immunohistochemistry

Journal: Journal of Inflammation Research

Article Title: Augmented Cornus officinalis Levels in Liuwei Dihuang Decoction Inhibits Nucleus Pulposus Cell Pyroptosis to Enhance Therapeutic Efficacy Against Intervertebral Disc Degeneration

doi: 10.2147/JIR.S465690

Figure Lengend Snippet: Combined Score of Modified Formulations of LWDHD in Delaying IVDD

Article Snippet: Primary antibodies against Tyrosine kinase A (TrkA), and NRF2 (S40) were obtained from HuaBio (Hangzhou, China).

Techniques: Modification, TUNEL Assay

Journal: Journal of Inflammation Research

Article Title: Augmented Cornus officinalis Levels in Liuwei Dihuang Decoction Inhibits Nucleus Pulposus Cell Pyroptosis to Enhance Therapeutic Efficacy Against Intervertebral Disc Degeneration

doi: 10.2147/JIR.S465690

Figure Lengend Snippet: Schematic illustration of the mechanism by which LWDHD formulation, containing varying dosages of CO, protects against IVDD progression through the inhibition of NP pyroptosis by activating the NRF2 pathway. The schematic illustration is drawn by Figdraw.

Article Snippet: Primary antibodies against Tyrosine kinase A (TrkA), and NRF2 (S40) were obtained from HuaBio (Hangzhou, China).

Techniques: Formulation, Inhibition

Journal: Molecules

Article Title: JND4135, a New Type II TRK Inhibitor, Overcomes TRK xDFG and Other Mutation Resistance In Vitro and In Vivo

doi: 10.3390/molecules27196500

Figure Lengend Snippet: JND4135 displays TRK kinase inhibitory activities against both wild-type (WT) and resistant mutation. ( A ) structure of JND4135; and representative graphs of kinase inhibitory activities of JND4135 against TRKA ( B ), TRKB ( C ), TRKC ( D ) and TRKA-G667C ( E ). All kinase inhibitory assays included at least two independent experiments.

Article Snippet: Primary antibodies against phospho-TRKA/TRKB/TRKC (ab197071) and CDK4 (ab199728) were purchased from Abcam.

Techniques: Mutagenesis