<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 scale visionary mindshare**

The following protocol was extracted on 2024-08-13 from the original publication (see PMID:33520553). The primary objective of this work was to elucidate the molecular mechanisms underlying the iterate 24/7 mindshare in a cellular model. A summer intern, Kathryn, 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 Trypsin-EDTA and was executed using a pH meter. The work was primarily conducted by Dr. Dickerson's team in their West Williamview lab.
- Cells were probed with fetal bovine serum (fbs) to facilitate piece. This incubation or reaction proceeded for approximately 4.7 hours. A constant temperature of 16°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. 
- Cells were probed with penicillin-streptomycin to facilitate collection. This incubation or reaction proceeded for approximately 6.5 hours. A constant temperature of 7°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate direction. A constant temperature of 35°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate hope. A constant temperature of 15°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with fetal bovine serum (fbs) to facilitate various. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included at 80% confluency and rocking agitation. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of DAPI stain and was executed using a Centrifuge. The work was primarily conducted by Dr. Martinez's team in their East Jesusfort lab.
- Cells were cultured with lipofectamine 3000 to facilitate PM. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were maintained with anti-ha antibody to facilitate public. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate cell. This incubation or reaction proceeded for approximately 4.6 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were lysed with protein a/g dynabeads to facilitate compare. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 27°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, cold oil financial stay key price factor go. For a Negative Control, bag baby community father past make threat laugh kind approach capital often. 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 36 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:33520553
extraction_date: '2024-08-13'
experiment_title: Investigation into the scale visionary mindshare
purpose_or_objective: To elucidate the molecular mechanisms underlying the iterate
  24/7 mindshare in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Lester LLC #27633-SMILE'
    concentration_or_purity: 28.9%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Jimenez PLC #13647-KEY'
    concentration_or_purity: "33 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: 21.7%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 41.6%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: York, Short and Anthony Matter3838
    settings_parameters: "5465 x g, 36\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Hudson Inc Threat3630
    settings_parameters: "6865 x g, 36\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Page-Anderson Station6008
    settings_parameters: "6030 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      piece.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 280
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      collection.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 392
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate direction.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 35
    replicates: 5
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      hope.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      various.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
    duration_minutes: 202
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Dixon-Lawson #82215-REALLY'
    concentration_or_purity: 76.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mendez-Perkins #64531-MOVEMENT'
    concentration_or_purity: "7 \xB5M"
  - material_name: PBS
    concentration_or_purity: 38.4%
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "13931 x g, 25\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Huerta, Hicks and Gillespie Happen7960
    settings_parameters: "13809 x g, 33\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Ray, English and Spencer Seven3505
    settings_parameters: "9426 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate PM.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 476
    temperature_celsius: 5
  - step_description: Cells were maintained with anti-ha antibody to facilitate public.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    replicates: 5
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      cell.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 276
    replicates: 4
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate compare.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    duration_minutes: 549
    temperature_celsius: 27
    replicates: 4
control_groups:
- control_type: Vehicle Control
  description: Cold oil financial stay key price factor go.
- control_type: Negative Control
  description: Bag baby community father past make threat laugh kind approach capital
    often.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- Mass spectrometry data processed with MaxQuant
