cyppa  (Alomone Labs)

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency attenuates NMDA-stimulated Ca 2+ loading in cultured cortical neurons. Cultures were challenged for 5 or 10 min with 30 μM NMDA, 10 μM CNQX, and 2 μM nimodipine in medium containing 45 CaCl 2 . CMP: counts per minute. * P

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: Efficacy of pCMV-U6-siRNA constructs. (A)RT-PCR analysis showed that a 495 bp product from PSD-93 was inhibited significantly by siP3, GAPDH mRNA was used as a loading Control. siP3 significantly decreased the mRNA of PSD93 by 80.2% of control.(B) Confirmation by western blot analysis of silencing PSD93 expression, The blots were reprobed with anti-GAPDH antibody to verify protein loading. (C) Relative protein level of PSD-93 after siRNA transfection, PSD93 decreased by 79.5% compared to control group when transfected with siP3.

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Construct, Polymerase Chain Reaction, Western Blot, Expressing, Transfection

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency or knockdown protects against NMDA-stimulated neurotoxicity. Cortical neurons were cultured from WT and PSD-93 KO mice and treated with NMDA (0, 10, 20, 30, 40, and 60 μM) in the presence of 10 μM CNQX and 2 μM nimodipine. (A) Neuronal viability was assessed by MTT assay. (B) Percentage of neuronal death was determined by propidium iodide and calcein AM staining. * P

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture, Mouse Assay, MTT Assay, Staining

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: MK-801 attenuates NMDA-induced neurotoxicity in cultured cortical neurons from WT but not from PSD-93 KO mice. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). * P

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture, Mouse Assay, Staining, MTT Assay

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency has no effect on non-NMDA receptor-triggered neurotoxicity in cultured cortical neurons. Cultured neurons were treated with kainate at the doses shown in the presence of 10 μM MK-801 and 2 μM nimodipine. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). n = 6 repeats.

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture, Staining, MTT Assay

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency decreases NMDA-stimulated increase in cGMP in cultured cortical neurons. (A) NMDA-induced neurotoxicity was NOS-dependent in cultured cortical neurons. Cultured neurons were treated with NMDA (30 μM or 60 μM) with or without L-NAME (10 μM or 30 μM). Neuronal viability was determined by MTT assay (top) and percentage of neuronal death by propidium iodide and calcein AM staining (bottom). * P

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture, MTT Assay, Staining

Journal: Neuroscience

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency significantly reduces synaptic expression of NR2A and NR2B in cultured cortical neurons. (A) Representative Western blots showing the levels of PSD-93, PSD-95, NR2A, and NR2B in the total soluble and synaptosomal fractions. (B) Statistical summary of the densitometric analysis expressed relative to WT mice after normalization to corresponding β-actin or N-cadherin. ** P

Article Snippet: Membranes were blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2A (1:200, Upstate/Chemicon, Temecula, CA), rabbit anti-NR2B (1:200, Upstate/Chemicon), mouse anti-PSD-95 (1:1,000; Upstate/Chemicon), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Expressing, Cell Culture, Western Blot, Mouse Assay

Journal: Cancers

Article Title: Orai1 Boosts SK3 Channel Activation

doi: 10.3390/cancers13246357

Figure Lengend Snippet: SK3–Orai1 interplay in LNCaP cells: Inset table and chemical structures represent the used agonist and antagonist of SK and Orai1 channels. ( A ) Cell viability of LNCaP cells after 24, 48, 72, and 96 h upon the treatment with SK3 channel agonist 10 μM Cyppa, 5 μM NS309, and antagonist 30 μM NS8593, 10 mM 4-AP detected via MTS assay; ( B ) Block diagram represents the cell proliferation of LNCaP cells after 96 h upon conditions described in ( A ). The Mann–Whitney test was used for statistical comparison considering differences statistically significant at p

Article Snippet: Inhibitors (NS8593 hydrochloride Cat #: N-195, 4-AP Cat #: A-115) and activators (NS-309 Cat #: N-180, 1-EBIO Cat #: E-150, Cyppa Cat #: C-110) were purchased from Alomone Labs (Jerusalem, Israel).

Techniques: MTS Assay, Blocking Assay, MANN-WHITNEY

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: Ethanol’s Effects on Transient Receptor Potential Channel Expression in Brain Microvascular Endothelial Cells

doi: 10.1007/s11481-018-9796-3

Figure Lengend Snippet: Endothelial cell barrier permeability following treatment with a) different concentrations of EtOH or b) the TRPM7 antagonists, NS8593, CyPPA, or SKA-31. * p

Article Snippet: For Trpm 7 inhibition, three different Trpm 7 Channel Blockers, NS 8593, CyPPA and SKA-31 (Alomone Labs, Jerusalem, Israel) were added to the inserts at 50 μM, together with a tracer Sodium Fluorescein (NaF) at 10 μg/mL, and incubated in a humidified cell culture incubator at 37 °C with 5% CO2 for 24 h. One hundred microliter samples from both the upper insert and the lower chamber were transferred to a clear 96-well plate.

Techniques: Permeability