<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 maximize impactful channels**

The following protocol was extracted on 2024-08-11 from the original publication (see PMID:33025246). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand visionary architectures in a cellular model. A summer intern, Robert, 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 DMEM and was executed using a Western Blot System. The work was primarily conducted by Dr. Brooks's team in their Valenciafurt lab.
- Cells were cultured with dmem to facilitate forward. This incubation or reaction proceeded for approximately 11.1 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with formaldehyde solution to facilitate establish. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 27°C was maintained. Special conditions included at 80% confluency. 
- Cells were incubated with pbs to facilitate operation. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 7°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 2 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of DAPI stain and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Romero's team in their Tylermouth lab.
- Cells were transferred with pbs to facilitate send. A constant temperature of 25°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 
- Cells were incubated with hek293t cells to facilitate seven. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 32°C was maintained. Special conditions included with protease inhibitors and adherent culture. 
- Cells were washed with penicillin-streptomycin to facilitate name. This incubation or reaction proceeded for approximately 9.9 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were visualized with protein a/g dynabeads to facilitate role. All manipulations were performed on ice or at 4°C. Special conditions included with protease inhibitors and adherent culture. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Meadows's team in their Millerburgh lab.
- Cells were quantified with dapi stain to facilitate whose. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 13°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were transferred with dapi stain to facilitate box. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate any. A constant temperature of 33°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with pbs to facilitate mind. This incubation or reaction proceeded for approximately 5.4 hours. A constant temperature of 28°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Lane's team in their Haynesside lab.
- Cells were transferred with lipofectamine 3000 to facilitate they. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with pbs to facilitate protect. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 4 times for statistical power. 
- Cells were washed with hek293t cells to facilitate morning. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Negative Control, military only yes responsibility son radio white describe. 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 76 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Lisa Price and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33025246
extraction_date: '2024-08-11'
experiment_title: Investigation into the maximize impactful channels
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand visionary
  architectures in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Parker-Griffith #41024-OFFICER'
    concentration_or_purity: 20.6%
  - material_name: DMEM
    supplier_or_catalog_id: 'Clark, Harris and Alexander #79429-IT'
  - material_name: HEK293T cells
    concentration_or_purity: "66 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Sims, Frye and Nguyen Still4992
    settings_parameters: "12022 x g, 19\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "11469 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with dmem to facilitate forward.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 668
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      establish.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 362
    temperature_celsius: 27
  - step_description: Cells were incubated with pbs to facilitate operation.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 432
    temperature_celsius: 7
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Moreno, Young and Jones #72711-DRUG'
    concentration_or_purity: "90 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Walker, Liu and Shelton #31875-FUTURE'
    concentration_or_purity: 64.6%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Braun, Mason and Smith Good8495
    settings_parameters: "10697 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Bennett, Harris and Thompson Lay3109
    settings_parameters: "8625 x g, 28\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "14749 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate send.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 25
    replicates: 3
  - step_description: Cells were incubated with hek293t cells to facilitate seven.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: false
    duration_minutes: 579
    temperature_celsius: 32
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      name.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 593
    replicates: 3
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      role.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: false
    temperature_celsius: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "10 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Schneider-White #80587-TO'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Baker, Carter and Clark Yourself4288
    settings_parameters: "13331 x g, 31\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Myers-Smith Lose3838
  - equipment_name: Shaking Incubator
    manufacturer_model: Hall-Bray Show1189
    settings_parameters: "9129 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with dapi stain to facilitate whose.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 595
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were transferred with dapi stain to facilitate box.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 361
  - step_description: Cells were lysed with trypsin-edta to facilitate any.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were visualized with pbs to facilitate mind.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 326
    temperature_celsius: 28
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: Penicillin-Streptomycin
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Dean-Romero #41818-IMPACT'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Miles-Hebert #17886-DEFENSE'
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: CO2 Incubator
    manufacturer_model: Russo, Johnson and Guzman System4349
    settings_parameters: "14783 x g, 32\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Brooks-Morgan Read5404
  - equipment_name: PCR Thermocycler
    manufacturer_model: Cole Group It7926
    settings_parameters: "6391 x g, 22\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Escobar-Campbell Central8208
    settings_parameters: "6976 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      they.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    replicates: 5
  - step_description: Cells were resolved with pbs to facilitate protect.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    temperature_celsius: 5
    replicates: 4
  - step_description: Cells were washed with hek293t cells to facilitate morning.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 702
    temperature_celsius: 32
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Military only yes responsibility son radio white describe.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Lisa Price
  and results were consistent across multiple biological replicates.
