RASFs were seeded into 24-well plates (density 2–4 × 104/well) Then cells were cultrued for 12, 24, 48 h in the presence of 0, 1, 10, 50 ng/ml of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), or lipopolysaccharide (LPS) respectively, or combined with 10μmol/l of PDTC (an inhibitor of nuclear transcription factor kappa B (NF-кB), Abcam, ab141406), PD98059 (an inhibitor of extracellular regulated protein kinases (ERK-1/2), Abcam, ab120234), or Static (an inhibitor of signal transducers and activators of transcription (STAT3), Abcam, ab120952), respectively. For the siRNA blocking assay, 0, 1, 10, 50 ng/ml of TNF-α with or without 100 nmol/l siRNA-E2F2 were added to seeded RASFs for 24 h (see below).
Ab141406
Manufactured by Abcam
Sourced in United Kingdom
Ab141406 is a primary antibody that recognizes human Synaptotagmin 1. It is a rabbit monoclonal antibody designed for use in various immunological techniques.
Automatically generated - may contain errors
4 protocols using ab141406
1
Modulation of RASF Activation by Cytokines
2
Endothelial Cell Culture and Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human dermal microvascular endothelial cells (HDMECs) were cultured in Endothelial Cell Medium (ECM), supplemented with fetal bovine serum (FBS, 5% as the final concentration)，Endothelial Cell Growth Supplement (ECGS, 1% as the final concentration) and P/S (Penicillin streptomycin) solution (1% as the final concentration, Scien Cell, USA). For some experiments, following a 1–2h recovery period after removal of the transfection mixture, the following inhibitors and activator (Abcam, UK) were added: the NF-κB inhibitor PDTC (ab141406, 150 μM for 24h) and the NF-κB activator Fusicoccin (ab145208, 12.5mM for 48h) [20 ,21 ].
3
Mouse Fracture Model Cytokine Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the fracture model was successfully established, the mice in Group E were intraperitoneally injected with TNF-α (0.625 mg/mL/kg/day for 3 days; ab141406; Abcam, Cambridge, UK); the mice in Group F were intrathecally injected with PDTC (0.5 μg/10 μL/day; Sigma, GF314) until the end of experimental observation. Mice in Group A, Group B, Group C and Group D were injected with the same dose of normal saline [29 (link),30 (link)].
4
Intrathecal SiRNA and PDTC Delivery in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The drugs were administrated intrathecally in rats under inhalational anesthesia. Rats were briefly anesthetized with 2% sevoflurane, the lower back of the rat was shaved and sterilized. Using a Hamilton microsyringe, the drug was injected between the L4 and L5 vertebrae. A sudden slight flick of the tail implicated that the needle has just entered into the subarachnoid space. The drug was injected slowly in 30 s. After injecton, the needle was held in place for a further 10 s to prevent outflow. Finally, the incision site was closed in layers.
A small interfering RNA (siRNA) was designed to target rat ANXA10 based on the genomic sequence (Gene Bank Accession: NM_001109110.1). The targeted nucleotide sequences were: 5′-TGCGAGAAGCCTACTGTTTACAATACAGC-3′. A scrambled sequence was designed as negative control (NC siRNA). The siRNA (3 μg/μl, dissolved in 10 μl PBS) was injected intrathecally. PDTC (ab141406, Abcam, 0.5 μg/10 μl), was dissolved in dimethyl sulfoxide (DMSO) for intrathecal injection.
A small interfering RNA (siRNA) was designed to target rat ANXA10 based on the genomic sequence (Gene Bank Accession: NM_001109110.1). The targeted nucleotide sequences were: 5′-TGCGAGAAGCCTACTGTTTACAATACAGC-3′. A scrambled sequence was designed as negative control (NC siRNA). The siRNA (3 μg/μl, dissolved in 10 μl PBS) was injected intrathecally. PDTC (ab141406, Abcam, 0.5 μg/10 μl), was dissolved in dimethyl sulfoxide (DMSO) for intrathecal injection.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!