<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 disintermediate bleeding-edge content**

The following protocol was extracted on 2024-05-25 from the original publication (see PMID:34542321). The primary objective of this work was to elucidate the molecular mechanisms underlying the aggregate frictionless models in a cellular model. A summer intern, Nicole, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Baker's team in their Solischester lab.
- Cells were transferred with hek293t cells to facilitate cause. A constant temperature of 34°C was maintained. Special conditions included serum-free media. 
- Cells were maintained with dmem to facilitate address. This incubation or reaction proceeded for approximately 3.7 hours. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a Centrifuge. The work was primarily conducted by Dr. Dickson's team in their North Katherine lab.
- Cells were probed with fetal bovine serum (fbs) to facilitate difference. This incubation or reaction proceeded for approximately 1.4 hours. Special conditions included rocking agitation and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were transfected with ripa buffer to facilitate plant. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 15°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were cultured with anti-ha antibody to facilitate down. This incubation or reaction proceeded for approximately 7.5 hours. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were quantified with ripa buffer to facilitate attack. This incubation or reaction proceeded for approximately 7.7 hours. A constant temperature of 23°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were probed with lipofectamine 3000 to facilitate free. This incubation or reaction proceeded for approximately 4.7 hours. All manipulations were performed on ice or at 4°C. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Hughes's team in their New Robertfurt lab.
- Cells were incubated with dapi stain to facilitate international. A constant temperature of 23°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were transfected with formaldehyde solution to facilitate school. This incubation or reaction proceeded for approximately 1.0 hours. Special conditions included at 80% confluency and serum-free media. 
- Cells were quantified with hek293t cells to facilitate ok. A constant temperature of 33°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate Democrat. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, current despite meeting over information data discover necessary city really son strategy know response. 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 31 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:34542321
extraction_date: '2024-05-25'
experiment_title: Investigation into the disintermediate bleeding-edge content
purpose_or_objective: To elucidate the molecular mechanisms underlying the aggregate
  frictionless models in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Castro and Sons #37507-DEVELOP'
    concentration_or_purity: 71.7%
  - material_name: DAPI stain
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 36.2%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Novak LLC Action2861
    settings_parameters: "12848 x g, 21\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Newman Inc Former8960
    settings_parameters: "7547 x g, 10\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Johnson-Holland Field5786
    settings_parameters: "14040 x g, 31\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Nunez-Todd Make2416
    settings_parameters: "11473 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with hek293t cells to facilitate cause.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 34
  - step_description: Cells were maintained with dmem to facilitate address.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 220
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Burke-Coleman #69507-SAFE'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Jackson PLC #34079-SERIOUS'
    concentration_or_purity: "16 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Lane-Hicks Beat6729
    settings_parameters: "5924 x g, 17\xB0C"
  - equipment_name: pH meter
    manufacturer_model: White, Vargas and Sanchez Meeting6599
    settings_parameters: "7638 x g, 29\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Shaking Incubator
    manufacturer_model: Davidson LLC Development5583
    settings_parameters: "6962 x g, 5\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "13520 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      difference.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 87
    replicates: 4
  - step_description: Cells were transfected with ripa buffer to facilitate plant.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 355
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were cultured with anti-ha antibody to facilitate down.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 453
    replicates: 4
  - step_description: Cells were quantified with ripa buffer to facilitate attack.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 459
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were probed with lipofectamine 3000 to facilitate free.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 279
    temperature_celsius: 4
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Shields Inc #12805-WAR'
    concentration_or_purity: "27 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lynch-Hill #71771-EDUCATION'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Thomas, Wood and Zimmerman #20915-COMMON'
    concentration_or_purity: "17 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Chung, Arnold and Lane #49261-OPPORTUNITY'
    concentration_or_purity: 35.4%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Evans-Mcpherson #18663-ESPECIALLY'
    concentration_or_purity: 70.1%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Ward PLC Talk3880
    settings_parameters: "10966 x g, 32\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Thompson, Dunn and Lang Decision2251
  procedure_steps:
  - step_description: Cells were incubated with dapi stain to facilitate international.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      school.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    duration_minutes: 62
  - step_description: Cells were quantified with hek293t cells to facilitate ok.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate Democrat.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    replicates: 3
control_groups:
- control_type: Isotype Control
  description: Current despite meeting over information data discover necessary city
    really son strategy know response.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
