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Clozapine dihydrochloride

Manufactured by Hello Bio
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Sourced in United Kingdom, United States

Clozapine dihydrochloride is a chemical compound used as a laboratory reagent. It is the dihydrochloride salt form of the antipsychotic drug clozapine. Clozapine dihydrochloride is typically used in research and analytical applications.

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Lab products found in correlation

Scopolamine hydrobromide, by Bio-Techne (2 mentions) Oxotremorine m, by Bio-Techne (2 mentions) Clozapine n oxide dihydrochloride, by Hello Bio (2 mentions)

6 protocols using clozapine dihydrochloride

1

Neuronal Activation Modulates Attentional Shifting

Cited 1 time
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Three weeks after surgery, behavioral testing on the attentional set-shifting task began. Prior to testing, the DREADDS were activated by clozapine (Gomez et al. 2017 ; Tan et al. 2020 (link)) either by systemic injection (Experiment 1) or intracranial infusion (Experiment 2).
In Experiment 1, clozapine dihydrochloride (Hello Bio) was dissolved in sterile saline. Twenty minutes before the test began, all rats received an intraperitoneal (I.P.) injection of clozapine dihydrochloride at a dose of 4 mg/kg (as salt).
In Experiment 2, clozapine dihydrochloride (Hello Bio) was dissolved in sterile saline at a dose of 1 μg/μL (as salt). Fifteen minutes prior to testing, rats were lightly restrained, the obturators removed, and 33-gauge stainless steel infusion cannulae (Plastic One) that projected 2.0 mm beyond the tip of the guide cannulae were inserted. Each pair of infusion cannula was connected to two 5 μL Hamilton syringes (Bonaduz) mounted on 2 infusions pumps (Harvard Apparatus Ltd). A volume of 0.25 μL per hemisphere was infused over 1 min. The infusion cannulae were left in situ for a further 1 min to allow absorption of the bolus. The infusion cannulae were then removed and the obturators replaced.
Bubb E.J., Aggleton J.P., O’Mara S.M, & Nelson A.J. (2020). Chemogenetics Reveal an Anterior Cingulate–Thalamic Pathway for Attending to Task-Relevant Information. Cerebral Cortex (New York, NY), 31(4), 2169-2186.
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2

Chemogenetic Modulation of Attention Shifting

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Three weeks post-surgery, behavioral testing on the attentional set-shifting task began.
Prior to testing, the DREADDS were activated by clozapine (Gomez et al., 2017; Tan et al., 2020) either by systemic injection (Experiment 1) or intracranial infusion (Experiment 2).
In Experiment 1, clozapine dihydrochloride (Hello Bio, Bristol, UK) was dissolved in sterile saline. Twenty minutes before the test began, all rats received an intraperitoneal (I.P.) injection of clozapine dihydrochloride at a dose of 4 mg/kg (as salt).
In Experiment 2, clozapine dihydrochloride (Hello Bio, Bristol, UK) was dissolved in sterile saline at a dose of 1 µg/µl (as salt). Fifteen minutes prior to testing, rats were lightly restrained, the obturators removed, and 33-gage stainless steel infusion cannulae (Plastic One, Roanoke, VA, USA) that projected 2.0 mm beyond the tip of the guide cannulae were inserted. Each pair of infusion cannula was connected to two 5 μl Hamilton syringes (Bonaduz, Switzerland) mounted on two infusions pumps (Harvard Apparatus Ltd, Kent, UK).
A volume of 0.25 μl per hemisphere was infused over 1 minute. The infusion cannulae were left in situ for a further 1 minute to allow absorption of the bolus. The infusion cannulae were then removed and the obturators replaced.
Bubb E.J., Aggleton J.P., O’Mara S.M., & Nelson A.J.D. (2020). Chemogenetics reveal an anterior cingulate-thalamic pathway for attending to task-relevant information.
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3

Rapid Neuronal Activation Mapping in Rats

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One week after the completion of behavioral testing, all rats received an I.P. injection of clozapine dihydrochloride (4 mg/kg as salt; HelloBio) to activate DREADDS in the experimental group. Each rat was placed individually in a cage in a dark holding room for 20 min, followed by 2 novel environments, each for 15 min. The first was a large square open field arena measuring 100 cm long, 100 cm wide, and 45 cm tall, which was filled with bedding sawdust and 6 novels objects. The second was a bow–tie maze (Albasser et al. 2010 (link)), which measured 120 cm long, 50 cm wide, and 50 cm tall, which was filled with bedding sawdust and food rewards (Cheerios, Nestle). Rats were then returned to the dark holding room for 90 min, to allow for neuronal Fos expression (Bisler et al. 2002 (link)) before perfusion.
Bubb E.J., Aggleton J.P., O’Mara S.M, & Nelson A.J. (2020). Chemogenetics Reveal an Anterior Cingulate–Thalamic Pathway for Attending to Task-Relevant Information. Cerebral Cortex (New York, NY), 31(4), 2169-2186.
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4

Pharmacological Agents for Neuromodulation

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Clozapine N-oxide (CNO) dihydrochloride (water soluble) and clozapine dihydrochloride (water soluble) were purchased from Hello Bio Inc. (Princeton, NJ, USA). Scopolamine hydrobromide, Oxotremorine M, and aCSF were purchased from Tocris (Minneapolis, MN, USA). Stock solutions of drugs were made and working concentrations of the drugs were prepared freshly on the day of each experiment by appropriate dilutions of the stock solutions in aCSF.
Yavas E., Trott J.M, & Fanselow M.S. (2021). Sexually dimorphic muscarinic acetylcholine receptor modulation of contextual fear learning in the dentate gyrus. Neurobiology of learning and memory, 185, 107528.
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5

Pharmacological Agents for Neuromodulation

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Clozapine N-oxide (CNO) dihydrochloride (water soluble) and clozapine dihydrochloride (water soluble) were purchased from Hello Bio Inc. (Princeton, NJ, USA). Scopolamine hydrobromide, Oxotremorine M, and aCSF were purchased from Tocris (Minneapolis, MN, USA). Stock solutions of drugs were made and working concentrations of the drugs were prepared freshly on the day of each experiment by appropriate dilutions of the stock solutions in aCSF.
Yavas E., Trott J.M, & Fanselow M.S. (2021). Sexually dimorphic muscarinic acetylcholine receptor modulation of contextual fear learning in the dentate gyrus. Neurobiology of learning and memory, 185, 107528.
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6

Investigating DREADDS-Induced Neuronal Activity

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One week after the completion of behavioral testing, the in-vivo influence of DREADDs was investigated by comparing group differences in the expression of the immediate-early gene c-fos. This gene is an indirect marker of neuronal activity (Chaudhuri, 1997; Tischmeyer and Grimm, 1999; Guzowski et al., 2005) and is expressed in multiple brain regions, including the anterior cingulate cortex and anterior thalamic nuclei following exposure to novelty (Zhu et al., 1995; Vann et al., 2000; Wirtshafter, 2005) .
Rats received an I.P. injection of clozapine dihydrochloride (4mg/kg as salt; HelloBio, Bristol, UK) to activate DREADDS and were placed in a cage in a dark holding room for 20 minutes.
Rats were then were placed in two novel environments, each for 15 minutes. The first was a large square open field arena measuring 100cm long, 100cm wide and 45cm tall which was filled with bedding sawdust and six novels objects. The second was a bow-tie maze (Albasser et al., 2010) which measured 120cm long, 50cm wide and 50cm tall and was filled with bedding sawdust and food rewards (Cheerios, Nestle, Glasgow, UK). Rats were then returned to the dark holding room for 90 minutes, to allow for neuronal Fos expression (Bisler et al., 2002) before perfusion.
Bubb E.J., Aggleton J.P., O’Mara S.M., & Nelson A.J.D. (2020). Chemogenetics reveal an anterior cingulate-thalamic pathway for attending to task-relevant information.
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