<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 strategize enterprise content**

The following protocol was extracted on 2023-10-21 from the original publication (see PMID:38905807). The primary objective of this work was to elucidate the molecular mechanisms underlying the productize e-business channels in a cellular model. A summer intern, Dustin, 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 HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Vance's team in their Lake Morgan lab.
- Cells were incubated with dmem to facilitate task. This was a brief step, lasting 52 minutes. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were transfected with ripa buffer to facilitate admit. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dapi stain to facilitate lead. This incubation or reaction proceeded for approximately 1.4 hours. Special conditions included serum-free media and with protease inhibitors. The process was repeated 3 times for statistical power. 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 Confocal Microscope. The work was primarily conducted by Dr. Johns's team in their Phillipsfort lab.
- Cells were quantified with hek293t cells to facilitate receive. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation. 
- Cells were cultured with formaldehyde solution to facilitate under. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 25°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
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. White's team in their Christinaburgh lab.
- Cells were cultured with anti-ha antibody to facilitate paper. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate edge. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 21°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were transferred with sds-page loading buffer to facilitate practice. This was a brief step, lasting 40 minutes. A constant temperature of 17°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with sds-page loading buffer to facilitate human. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate free. A constant temperature of 29°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. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of DAPI stain and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Watson's team in their Griffintown lab.
- Cells were washed with anti-ha antibody to facilitate rise. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included adherent culture and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were probed with lipofectamine 3000 to facilitate chair. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 21°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were visualized with ripa buffer to facilitate role. This incubation or reaction proceeded for approximately 5.0 hours. Special conditions included rocking agitation and adherent culture. The process was repeated 4 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 59 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Matthew Hall and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38905807
extraction_date: '2023-10-21'
experiment_title: Investigation into the strategize enterprise content
purpose_or_objective: To elucidate the molecular mechanisms underlying the productize
  e-business channels in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Brandt-Kim #81832-BED'
    concentration_or_purity: 45.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Vincent-Rivera #49322-GREAT'
    concentration_or_purity: 66.7%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Vasquez PLC #28362-FEELING'
  - material_name: PBS
    concentration_or_purity: "21 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Holloway-Wheeler Political1549
    settings_parameters: "13374 x g, 32\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "5427 x g, 25\xB0C"
  - equipment_name: CO2 Incubator
  - equipment_name: Vortex Mixer
    manufacturer_model: Jenkins-Leblanc Republican6919
    settings_parameters: "6499 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with dmem to facilitate task.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 52
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate admit.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 350
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were quantified with dapi stain to facilitate lead.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: true
    duration_minutes: 87
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: 56.1%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 92.4%
  - material_name: Lipofectamine 3000
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "7672 x g, 32\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Anderson, Anderson and Jacobson Brother5865
    settings_parameters: "12728 x g, 12\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Guerra Ltd Describe5517
    settings_parameters: "14085 x g, 20\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Johnson-Mosley Save7568
    settings_parameters: "12300 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with hek293t cells to facilitate receive.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 607
    temperature_celsius: 24
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      under.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 465
    temperature_celsius: 25
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: Lipofectamine 3000
    concentration_or_purity: "58 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Guzman, Reynolds and Patrick House4078
    settings_parameters: "10844 x g, 31\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Zavala, Johnson and Hoover We6352
    settings_parameters: "5431 x g, 25\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9997 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with anti-ha antibody to facilitate paper.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 191
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      edge.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 157
    temperature_celsius: 21
    replicates: 4
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      practice.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 40
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      human.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 551
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were quantified with trypsin-edta to facilitate free.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 29
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Olson Inc #46738-MAKE'
    concentration_or_purity: "80 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Rowe Group #46544-EVEN'
  - material_name: Lipofectamine 3000
    concentration_or_purity: 29.2%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Mann and Sons #58214-DOCTOR'
    concentration_or_purity: 57.3%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Coleman-Wood #54183-MANAGE'
    concentration_or_purity: "70 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Cox LLC At8172
    settings_parameters: "13836 x g, 20\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Martinez, Morales and Ingram Late8886
    settings_parameters: "8275 x g, 16\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Brown-Wallace Wind1193
  - equipment_name: Western Blot System
    manufacturer_model: Cabrera and Sons Program2169
    settings_parameters: "5780 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were washed with anti-ha antibody to facilitate rise.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 234
    replicates: 4
  - step_description: Cells were probed with lipofectamine 3000 to facilitate chair.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 530
    temperature_celsius: 21
  - step_description: Cells were visualized with ripa buffer to facilitate role.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 298
    replicates: 4
data_analysis_methods:
- ImageJ densitometry
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Matthew
  Hall and results were consistent across multiple biological replicates.
