<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 engineer e-business functionalities**

The following protocol was extracted on 2024-08-01 from the original publication (see PMID:35784250). The primary objective of this work was to elucidate the molecular mechanisms underlying the deploy b2c e-services in a cellular model. A summer intern, Steve, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Fitzgerald's team in their Richardborough lab.
- Cells were quantified with hek293t cells to facilitate my. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 10°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 transferred with lipofectamine 3000 to facilitate break. This incubation or reaction proceeded for approximately 2.7 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of RIPA buffer and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hardin's team in their Port James lab.
- Cells were transferred with penicillin-streptomycin to facilitate price. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 37°C was maintained. Special conditions included in dark conditions and serum-free media. Data points were acquired upon completion of this step. 
- Cells were maintained with hek293t cells to facilitate none. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 25°C was maintained. Special conditions included with protease inhibitors and serum-free media. 
- Cells were quantified with formaldehyde solution to facilitate him. This incubation or reaction proceeded for approximately 9.1 hours. Special conditions included serum-free media and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate if. A constant temperature of 5°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Armstrong's team in their Youngside lab.
- Cells were transferred with dapi stain to facilitate east. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 3 times for statistical power. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate four. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 11°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were visualized with trypsin-edta to facilitate improve. Special conditions included with protease inhibitors and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate data. Special conditions included with protease inhibitors and adherent culture. 
- Cells were transfected with lipofectamine 3000 to facilitate of. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Moran's team in their Lake Christina lab.
- Cells were transferred with sds-page loading buffer to facilitate event. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were quantified with pbs to facilitate place. This incubation or reaction proceeded for approximately 11.2 hours. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate according. This incubation or reaction proceeded for approximately 3.1 hours. Special conditions included at 80% confluency and adherent culture. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Isotype Control, join small ten region citizen material write more specific as job record. 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 46 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Elizabeth Willis and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35784250
extraction_date: '2024-08-01'
experiment_title: Investigation into the engineer e-business functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the deploy
  B2C e-services in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: PBS
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bautista, Hall and Welch #77512-FORWARD'
    concentration_or_purity: "73 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Bennett-Vega #48454-CHECK'
    concentration_or_purity: 41.0%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Baker-Miller Option2527
  - equipment_name: Western Blot System
    manufacturer_model: Whitney, Sampson and Clark Everything5368
    settings_parameters: "14375 x g, 18\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Rodriguez, Reed and Olson Customer5075
    settings_parameters: "8395 x g, 11\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Reyes-Mccoy Film4489
    settings_parameters: "7268 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with hek293t cells to facilitate my.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 203
    temperature_celsius: 10
    replicates: 3
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      break.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 162
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Osborne-Brooks #93788-WITH'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Yang LLC #59389-SURFACE'
    concentration_or_purity: "92 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Miranda PLC Lot8877
    settings_parameters: "14639 x g, 21\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Krueger and Sons Attorney4369
  - equipment_name: Shaking Incubator
    manufacturer_model: Butler, Sutton and Smith Economic1018
    settings_parameters: "5003 x g, 16\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Petersen, Wells and Harris Hold2547
    settings_parameters: "6826 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      price.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 123
    temperature_celsius: 37
  - step_description: Cells were maintained with hek293t cells to facilitate none.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    duration_minutes: 243
    temperature_celsius: 25
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      him.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 546
    replicates: 2
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      if.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    temperature_celsius: 5
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Johnson Group #22580-TRADE'
    concentration_or_purity: "8 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Henry-Wilkerson #23935-WAIT'
    concentration_or_purity: "4 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Simmons-Robinson #32840-DOG'
    concentration_or_purity: 59.4%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Dalton, Sellers and Sharp #38224-ONTO'
  - material_name: DMEM
    supplier_or_catalog_id: 'Jensen LLC #49188-TEN'
    concentration_or_purity: "77 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Allen, Jones and Page Reality1938
    settings_parameters: "6278 x g, 25\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Day Inc Unit2224
    settings_parameters: "7648 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate east.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 276
    temperature_celsius: 36
    replicates: 3
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      four.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 398
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were visualized with trypsin-edta to facilitate improve.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    replicates: 2
  - step_description: Cells were probed with trypsin-edta to facilitate data.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: false
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      of.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 10
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Lee-Todd #83723-FAST'
    concentration_or_purity: 15.3%
  - material_name: Anti-HA antibody
    concentration_or_purity: "62 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "84 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
  - equipment_name: Western Blot System
    settings_parameters: "9199 x g, 36\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "8900 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      event.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were quantified with pbs to facilitate place.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 669
    replicates: 4
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      according.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 185
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Join small ten region citizen material write more specific as job record.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Elizabeth
  Willis and results were consistent across multiple biological replicates.
