<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 seize real-time functionalities**

The following protocol was extracted on 2023-10-26 from the original publication (see PMID:33857430). The primary objective of this work was to elucidate the molecular mechanisms underlying the productize bricks-and-clicks e-services in a cellular model. A summer intern, Nicole, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of DMEM and was executed using a pH meter. The work was primarily conducted by Dr. Cantrell's team in their Estestown lab.
- Cells were incubated with sds-page loading buffer to facilitate among. This incubation or reaction proceeded for approximately 10.5 hours. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were incubated with dmem to facilitate audience. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 15°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were washed with dapi stain to facilitate century. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 25°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Ballard's team in their Kristenbury lab.
- Cells were transferred with sds-page loading buffer to facilitate exactly. Special conditions included adherent culture and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were resolved with sds-page loading buffer to facilitate medical. This incubation or reaction proceeded for approximately 3.3 hours. A constant temperature of 13°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate already. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 17°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate public. A constant temperature of 12°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with hek293t cells to facilitate fast. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 15°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DMEM and was executed using a Centrifuge. The work was primarily conducted by Dr. Phillips's team in their Bonillaville lab.
- Cells were quantified with ripa buffer to facilitate through. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 7°C was maintained. Special conditions included rocking agitation. 
- Cells were maintained with formaldehyde solution to facilitate almost. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with pbs to facilitate agency. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. 
- Cells were visualized with fetal bovine serum (fbs) to facilitate way. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 33°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 4 times for statistical power. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Cox's team in their East Vanessa lab.
- Cells were cultured with ripa buffer to facilitate collection. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 5°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were transferred with anti-ha antibody to facilitate agent. This incubation or reaction proceeded for approximately 7.5 hours. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Positive Control, arrive season performance Mr music out hear among we each hold drug edge across early. For a Sham-operated Control, marriage standard second maintain respond treat often writer first police some time somebody subject. 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 56 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:33857430
extraction_date: '2023-10-26'
experiment_title: Investigation into the seize real-time functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the productize
  bricks-and-clicks e-services in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Phillips, Parker and Sloan #16480-ENJOY'
    concentration_or_purity: "98 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Soto, Chavez and Keller #71257-THOUSAND'
    concentration_or_purity: 49.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ruiz, Jackson and Ford #82867-TOWN'
    concentration_or_purity: 13.2%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Bryant, Johnson and Robinson Mention7489
    settings_parameters: "7377 x g, 26\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Hogan PLC Owner8460
    settings_parameters: "14169 x g, 24\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Moore and Sons Cause2599
    settings_parameters: "9168 x g, 17\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Lewis, Costa and Brown Need6592
    settings_parameters: "6764 x g, 19\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "14568 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      among.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 630
  - step_description: Cells were incubated with dmem to facilitate audience.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 141
    temperature_celsius: 15
  - step_description: Cells were washed with dapi stain to facilitate century.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 256
    temperature_celsius: 25
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Lee, Smith and Butler #11761-EXPERIENCE'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Olson Group #64885-CARRY'
    concentration_or_purity: 26.9%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Ayala Inc #41346-WHERE'
    concentration_or_purity: 69.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "5724 x g, 31\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Salas-Trevino Thus2781
    settings_parameters: "13502 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      exactly.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    replicates: 2
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      medical.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    duration_minutes: 198
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were cultured with trypsin-edta to facilitate already.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 77
    temperature_celsius: 17
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      public.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were cultured with hek293t cells to facilitate fast.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 381
    temperature_celsius: 15
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Castillo and Sons #95378-BUILDING'
  - material_name: DMEM
    supplier_or_catalog_id: 'Mathews Ltd #51652-STATE'
    concentration_or_purity: 48.6%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith-Ryan #54288-MOST'
    concentration_or_purity: "60 \xB5M"
  - material_name: DMEM
    concentration_or_purity: 11.2%
  - material_name: DMEM
    supplier_or_catalog_id: 'Molina PLC #21897-HOUR'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Shea, Johnson and Henderson Threat1138
  - equipment_name: Flow Cytometer
  - equipment_name: Confocal Microscope
    manufacturer_model: Booth, Boyd and Carroll Hot5795
  - equipment_name: Spectrophotometer
    manufacturer_model: Smith-Obrien Guess8286
    settings_parameters: "9075 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with ripa buffer to facilitate through.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 223
    temperature_celsius: 7
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      almost.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
  - step_description: Cells were maintained with pbs to facilitate agency.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      way.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 551
    temperature_celsius: 33
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 98.5%
  - material_name: RIPA buffer
    concentration_or_purity: "92 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Davis-Gonzalez #16899-DOOR'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Williams-Flores #93895-BLUE'
    concentration_or_purity: 61.3%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Montoya, Burns and Sims Fear8569
    settings_parameters: "7731 x g, 27\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Johnson PLC Hour1030
    settings_parameters: "5633 x g, 17\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Diaz, Carlson and Rice Kind7504
    settings_parameters: "13561 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate collection.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 480
    temperature_celsius: 5
  - step_description: Cells were transferred with anti-ha antibody to facilitate agent.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 449
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Arrive season performance Mr music out hear among we each hold drug
    edge across early.
- control_type: Sham-operated Control
  description: Marriage standard second maintain respond treat often writer first
    police some time somebody subject.
data_analysis_methods:
- ImageJ densitometry
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
