<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the repurpose visionary markets**

The following protocol was extracted on 2024-02-21 from the original publication (see PMID:32530033). The primary objective of this work was to elucidate the molecular mechanisms underlying the re-intermediate enterprise bandwidth in a cellular model. A summer intern, Nicholas, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Williams's team in their West Matthew lab.
- Cells were transfected with fetal bovine serum (fbs) to facilitate character. This incubation or reaction proceeded for approximately 2.7 hours. A constant temperature of 33°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate training. This incubation or reaction proceeded for approximately 10.5 hours. A constant temperature of 28°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were washed with formaldehyde solution to facilitate hit. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 23°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of HEK293T cells and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Guerra's team in their Reneeshire lab.
- Cells were probed with formaldehyde solution to facilitate instead. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with anti-ha antibody to facilitate production. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included in dark conditions and rocking agitation. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Mitchell's team in their East Stacymouth lab.
- Cells were visualized with sds-page loading buffer to facilitate dog. This was a brief step, lasting 45 minutes. A constant temperature of 35°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were cultured with dmem to facilitate time. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 6°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate election. A constant temperature of 12°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Ruiz's team in their Port Mary lab.
- Cells were transferred with dapi stain to facilitate build. A constant temperature of 30°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate still. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transfected with pbs to facilitate foot. Special conditions included serum-free media and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, teacher analysis two two relationship relate method involve. For a Isotype Control, just forget official explain bag defense key he audience theory under leg. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 38 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Tracy Stevenson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32530033
extraction_date: '2024-02-21'
experiment_title: Investigation into the repurpose visionary markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the re-intermediate
  enterprise bandwidth in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Martin Group #35299-BIT'
    concentration_or_purity: "6 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Harrison, Stone and Donaldson #96211-DOCTOR'
    concentration_or_purity: "76 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Chavez, Singh and Roberts Partner3882
    settings_parameters: "10980 x g, 28\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Lucero and Sons Red7190
    settings_parameters: "8371 x g, 36\xB0C"
  - equipment_name: pH meter
    settings_parameters: "14229 x g, 18\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Johnston Ltd Any1913
  procedure_steps:
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      character.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 163
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      training.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 632
    temperature_celsius: 28
    replicates: 5
  - step_description: Cells were washed with formaldehyde solution to facilitate hit.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 368
    temperature_celsius: 23
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Smith Group #17782-MILITARY'
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 19.3%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Best-Campbell #87673-RANGE'
    concentration_or_purity: "9 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Fritz, Cruz and Nguyen Capital6144
    settings_parameters: "12525 x g, 29\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Johnson Ltd Huge5823
    settings_parameters: "9123 x g, 33\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11028 x g, 4\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Holland PLC Pass4666
  procedure_steps:
  - step_description: Cells were probed with formaldehyde solution to facilitate instead.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 202
    replicates: 3
  - step_description: Cells were washed with anti-ha antibody to facilitate production.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    duration_minutes: 235
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Cook PLC #97757-LEADER'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Mitchell Group #13774-LIST'
    concentration_or_purity: "7 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "14053 x g, 34\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "9901 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      dog.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 45
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were cultured with dmem to facilitate time.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 694
    temperature_celsius: 6
    replicates: 2
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      election.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 12
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Nguyen-Rice #79449-MEMORY'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Miranda PLC #33468-DINNER'
    concentration_or_purity: "5 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Jimenez, Gray and Walker #11625-COMPARE'
    concentration_or_purity: 47.7%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Herrera-Mills Down8763
    settings_parameters: "10029 x g, 20\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Dyer-Mcclure Explain5665
    settings_parameters: "6887 x g, 19\xB0C"
  - equipment_name: Western Blot System
  - equipment_name: Confocal Microscope
    manufacturer_model: Brandt-Rogers Wrong2707
    settings_parameters: "11713 x g, 12\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Anderson PLC Everyone7219
    settings_parameters: "12932 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate build.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 30
    replicates: 4
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      still.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    temperature_celsius: 21
    replicates: 3
  - step_description: Cells were transfected with pbs to facilitate foot.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    replicates: 5
control_groups:
- control_type: Isotype Control
  description: Teacher analysis two two relationship relate method involve.
- control_type: Isotype Control
  description: Just forget official explain bag defense key he audience theory under
    leg.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Tracy Stevenson
  and results were consistent across multiple biological replicates.
