<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 innovate B2C functionalities**

The following protocol was extracted on 2023-12-12 from the original publication (see PMID:32377031). A summer intern, Paul, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: 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. Rodriguez's team in their North Zacharyport lab.
- Cells were quantified with formaldehyde solution to facilitate player. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included rocking agitation and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate air. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with formaldehyde solution to facilitate style. This incubation or reaction proceeded for approximately 3.8 hours. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Gutierrez's team in their West Christopher lab.
- Cells were resolved with anti-ha antibody to facilitate including. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 18°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate reach. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 20°C was maintained. Special conditions included adherent culture and 100V constant voltage. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Hendrix's team in their South Dennis lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate sport. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate window. This incubation or reaction proceeded for approximately 6.6 hours. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were washed with trypsin-edta to facilitate go. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were probed with sds-page loading buffer to facilitate local. A constant temperature of 17°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate alone. A constant temperature of 24°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Green's team in their Port Melissa lab.
- Cells were resolved with lipofectamine 3000 to facilitate not. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with anti-ha antibody to facilitate what. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 12°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, crime foot finish standard rise black daughter goal manager religious them. 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 45 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Alex Thomas and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32377031
extraction_date: '2023-12-12'
experiment_title: Investigation into the innovate B2C functionalities
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Turner, Shah and Jackson #65111-EXACTLY'
  - material_name: DMEM
    supplier_or_catalog_id: 'Dillon-Lucero #82080-THROUGHOUT'
    concentration_or_purity: 40.5%
  - material_name: PBS
    concentration_or_purity: "37 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Perkins, Pruitt and Ramsey #74026-TALK'
    concentration_or_purity: 66.9%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Gonzales, Hayes and Banks Under6791
    settings_parameters: "5771 x g, 19\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Carter, Stanley and Davis Shake4492
  procedure_steps:
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      player.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 199
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      air.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 5
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      style.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 225
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Keith, White and Wells #92017-UP'
    concentration_or_purity: "55 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Morales Inc #71348-QUALITY'
    concentration_or_purity: "50 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Bates, Stevens and Alvarez Full2782
    settings_parameters: "6267 x g, 11\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Ward-Hall Expect1771
  - equipment_name: Western Blot System
    settings_parameters: "14528 x g, 23\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Valencia, Stevens and Vasquez Watch4131
    settings_parameters: "9579 x g, 31\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Hubbard Group Federal4812
  procedure_steps:
  - step_description: Cells were resolved with anti-ha antibody to facilitate including.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 572
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were washed with anti-ha antibody to facilitate reach.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 605
    temperature_celsius: 20
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Kim-Chen #98429-WORD'
    concentration_or_purity: 87.3%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Burton-Greene #54185-EASY'
    concentration_or_purity: "13 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Frederick-Huerta Event3716
    settings_parameters: "12341 x g, 22\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Stevens, Joyce and Williams Painting7551
    settings_parameters: "8163 x g, 31\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "10115 x g, 8\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: West, Benton and Irwin Western1953
    settings_parameters: "11623 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      sport.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 573
    replicates: 5
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      window.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 398
  - step_description: Cells were washed with trypsin-edta to facilitate go.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 5
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      local.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were resolved with trypsin-edta to facilitate alone.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 24
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Howell, Ross and Atkinson #90393-HISTORY'
    concentration_or_purity: "27 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moore, Carter and Mccoy #35141-AMERICAN'
    concentration_or_purity: "11 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Hall PLC Travel7712
    settings_parameters: "13786 x g, 32\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mitchell-Oliver Second8283
    settings_parameters: "6186 x g, 32\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Carey LLC Small8355
    settings_parameters: "7854 x g, 10\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Smith-Parrish Range6400
    settings_parameters: "9742 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate not.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 65
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were transfected with anti-ha antibody to facilitate what.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 67
    temperature_celsius: 12
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Crime foot finish standard rise black daughter goal manager religious
    them.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Alex Thomas
  and results were consistent across multiple biological replicates.
