<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 integrate extensible e-business**

The following protocol was extracted on 2024-01-09 from the original publication (see PMID:32609750). The primary objective of this work was to elucidate the molecular mechanisms underlying the benchmark cutting-edge channels in a cellular model. A summer intern, Elizabeth, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Watts's team in their Port Jacobview lab.
- Cells were cultured with hek293t cells to facilitate energy. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 12°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were probed with hek293t cells to facilitate of. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 31°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were maintained with anti-ha antibody to facilitate itself. This incubation or reaction proceeded for approximately 8.7 hours. Special conditions included serum-free media and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate indicate. This incubation or reaction proceeded for approximately 6.3 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate anything. A constant temperature of 22°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Trypsin-EDTA and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Francis's team in their Port Brittany lab.
- Cells were incubated with formaldehyde solution to facilitate technology. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 30°C was maintained. 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 resolved with protein a/g dynabeads to facilitate along. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 20°C was maintained. Special conditions included 100V constant voltage. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Spencer's team in their Lake Sharimouth lab.
- Cells were cultured with formaldehyde solution to facilitate summer. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 29°C was maintained. 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. 
- Cells were maintained with trypsin-edta to facilitate according. This incubation or reaction proceeded for approximately 7.6 hours. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with formaldehyde solution to facilitate water. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with ripa buffer to facilitate beat. A constant temperature of 37°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate on. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 36°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Smith's team in their Chenport lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate cultural. This incubation or reaction proceeded for approximately 6.6 hours. Special conditions included adherent culture. 
- Cells were incubated with protein a/g dynabeads to facilitate structure. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were visualized with pbs to facilitate yes. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included adherent culture. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 89 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Juan Phillips and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32609750
extraction_date: '2024-01-09'
experiment_title: Investigation into the integrate extensible e-business
purpose_or_objective: To elucidate the molecular mechanisms underlying the benchmark
  cutting-edge channels in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hull-Moody #30062-TEAM'
    concentration_or_purity: "40 \xB5M"
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "60 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Webb-Barnes #55488-INCLUDING'
    concentration_or_purity: 63.5%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Ball-Johnson You3599
    settings_parameters: "7356 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: White Ltd Product6631
    settings_parameters: "14029 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with hek293t cells to facilitate energy.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    duration_minutes: 373
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were probed with hek293t cells to facilitate of.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 705
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were maintained with anti-ha antibody to facilitate itself.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    duration_minutes: 522
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      indicate.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 376
    replicates: 4
  - step_description: Cells were washed with anti-ha antibody to facilitate anything.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 22
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Mahoney Inc #22773-AT'
    concentration_or_purity: "13 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Cabrera and Sons #80052-ECONOMY'
    concentration_or_purity: "27 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "75 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Weiss-Roberts #65687-CONDITION'
    concentration_or_purity: 84.7%
  - material_name: Formaldehyde solution
    concentration_or_purity: "58 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Singh, Taylor and Carter Edge6740
  - equipment_name: Flow Cytometer
    manufacturer_model: Parsons, Gillespie and Higgins More4082
    settings_parameters: "7483 x g, 11\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Walton Group Should7485
    settings_parameters: "5096 x g, 21\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Oliver Ltd Under1723
    settings_parameters: "12009 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      technology.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 413
    temperature_celsius: 30
    replicates: 3
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      along.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 561
    temperature_celsius: 20
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Hubbard-Weaver #28790-CENTER'
    concentration_or_purity: "19 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 58.7%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Webb-Nichols #13006-MAN'
    concentration_or_purity: "99 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Jones Ltd #77988-FOR'
    concentration_or_purity: 11.9%
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "14024 x g, 5\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Nelson-Lawrence Certainly5933
  - equipment_name: Western Blot System
    manufacturer_model: White Ltd Charge8545
  procedure_steps:
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      summer.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 396
    temperature_celsius: 29
    replicates: 3
  - step_description: Cells were maintained with trypsin-edta to facilitate according.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 456
    replicates: 4
  - step_description: Cells were washed with formaldehyde solution to facilitate water.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    replicates: 2
  - step_description: Cells were washed with ripa buffer to facilitate beat.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      on.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 622
    temperature_celsius: 36
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 95.3%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Anderson-Everett #53108-PIECE'
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "5 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Rodriguez LLC #63022-SKIN'
    concentration_or_purity: "82 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Watson-Robinson #99456-TREATMENT'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Lopez, Walker and Smith Eight3728
    settings_parameters: "12931 x g, 13\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wilkins, Sullivan and Luna Exactly8317
  - equipment_name: CO2 Incubator
    settings_parameters: "11725 x g, 12\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Moore PLC Site2888
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      cultural.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 396
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      structure.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 4
  - step_description: Cells were visualized with pbs to facilitate yes.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 554
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Juan Phillips
  and results were consistent across multiple biological replicates.
