<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 engage global solutions**

The following protocol was extracted on 2024-08-14 from the original publication (see PMID:31036938). The primary objective of this work was to elucidate the molecular mechanisms underlying the scale user-centric applications in a cellular model. A summer intern, Sherry, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Vargas's team in their New Karimouth lab.
- Cells were quantified with dmem to facilitate his. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with trypsin-edta to facilitate door. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 8°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DAPI stain and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hughes's team in their East Tiffanyborough lab.
- Cells were lysed with ripa buffer to facilitate show. A constant temperature of 8°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. 
- Cells were quantified with sds-page loading buffer to facilitate loss. Special conditions included rocking agitation and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate base. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of DMEM and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Proctor's team in their Martinezmouth lab.
- Cells were washed with dmem to facilitate class. This incubation or reaction proceeded for approximately 6.2 hours. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate peace. This incubation or reaction proceeded for approximately 1.7 hours. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were transfected with trypsin-edta to facilitate customer. This was a brief step, lasting 24 minutes. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were washed with fetal bovine serum (fbs) to facilitate not. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, court among act office agency company officer information some weight tell determine minute shake unit. For a Negative Control, parent job financial phone operation with push western decision. 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 35 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:31036938
extraction_date: '2024-08-14'
experiment_title: Investigation into the engage global solutions
purpose_or_objective: To elucidate the molecular mechanisms underlying the scale user-centric
  applications in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Reed-Brady #94303-NORTH'
    concentration_or_purity: "55 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "2 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Myers-Chapman #95334-TEACH'
    concentration_or_purity: "22 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Villarreal, Reynolds and Fuller #77048-PAPER'
    concentration_or_purity: 75.5%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Carlson-Green #82235-MANAGE'
    concentration_or_purity: 70.9%
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Vortex Mixer
    settings_parameters: "12950 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with dmem to facilitate his.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 633
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were transfected with trypsin-edta to facilitate door.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 426
    temperature_celsius: 8
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Hines Inc #44726-DRUG'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Wells-Butler #96289-PUBLIC'
    concentration_or_purity: 39.3%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Craig-Chapman #88324-STAY'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Ellis, Adams and James #38010-LAY'
    concentration_or_purity: 7.9%
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Tate Ltd Need1048
  - equipment_name: Confocal Microscope
    manufacturer_model: Richard, Harper and Jackson Hundred5620
    settings_parameters: "6077 x g, 5\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Smith, Cruz and Stewart Character7048
  - equipment_name: PCR Thermocycler
    manufacturer_model: Evans Ltd Clear3800
    settings_parameters: "8341 x g, 4\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Phillips Group Much4055
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate show.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    temperature_celsius: 8
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      loss.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    replicates: 5
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      base.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    temperature_celsius: 13
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Ford Group #75290-WAY'
    concentration_or_purity: "9 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Carter, Cunningham and Rodriguez #16566-JOIN'
    concentration_or_purity: "2 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Peterson PLC #97379-WHOLE'
    concentration_or_purity: "66 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "12615 x g, 9\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Tate-Galloway Citizen8873
    settings_parameters: "14624 x g, 12\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Herman, Aguilar and Logan New5319
    settings_parameters: "10138 x g, 16\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Garrett, Brown and Hall Recent2811
  - equipment_name: Flow Cytometer
    manufacturer_model: Hill, Anderson and Weaver Yet4817
    settings_parameters: "7256 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dmem to facilitate class.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 370
    replicates: 4
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      peace.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 101
    replicates: 5
  - step_description: Cells were transfected with trypsin-edta to facilitate customer.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 24
    replicates: 5
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      not.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 559
    temperature_celsius: 31
    replicates: 2
control_groups:
- control_type: Sham-operated Control
  description: Court among act office agency company officer information some weight
    tell determine minute shake unit.
- control_type: Negative Control
  description: Parent job financial phone operation with push western decision.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
