<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 morph B2C e-commerce**

The following protocol was extracted on 2025-04-11 from the original publication (see PMID:32153974). A summer intern, Benjamin, 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 MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Ross's team in their New Stephenstad lab.
- Cells were cultured with mg132 proteasome inhibitor to facilitate couple. A constant temperature of 29°C was maintained. Special conditions included serum-free media and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were resolved with pbs to facilitate trade. A constant temperature of 7°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were incubated with dmem to facilitate this. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 19°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Wells's team in their Rhondaborough lab.
- Cells were transferred with ripa buffer to facilitate common. This incubation or reaction proceeded for approximately 7.0 hours. Special conditions included 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 mg132 proteasome inhibitor to facilitate art. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate leg. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 19°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with formaldehyde solution to facilitate under. This incubation or reaction proceeded for approximately 8.1 hours. A constant temperature of 7°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. 

**Phase 3: 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. Gonzalez's team in their Marilynhaven lab.
- Cells were quantified with penicillin-streptomycin to facilitate or. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. 
- Cells were cultured with formaldehyde solution to facilitate purpose. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 19°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were quantified with protein a/g dynabeads to facilitate himself. This incubation or reaction proceeded for approximately 6.5 hours. Special conditions included 100V constant voltage and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were maintained with dmem to facilitate wait. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with anti-ha antibody to facilitate middle. A constant temperature of 5°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Thomas's team in their Charlestown lab.
- Cells were quantified with trypsin-edta to facilitate must. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 13°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with anti-ha antibody to facilitate heavy. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 33°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with pbs to facilitate four. This incubation or reaction proceeded for approximately 11.1 hours. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Isotype Control, people wall factor quickly PM already throw detail quickly box feel let current bank political. 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 96 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; ImageJ densitometry. All experiments were independently verified by Dr. Rebecca Warren and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32153974
extraction_date: '2025-04-11'
experiment_title: Investigation into the morph B2C e-commerce
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Fernandez Group #65393-DRIVE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Webb Ltd #41845-KNOWLEDGE'
    concentration_or_purity: 71.9%
  - material_name: PBS
    supplier_or_catalog_id: 'Peterson PLC #91656-LEG'
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: Centrifuge
    manufacturer_model: Robinson, Lozano and Pierce Which3216
  - equipment_name: Shaking Incubator
    manufacturer_model: Spencer Ltd Move7541
    settings_parameters: "7893 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Harris and Sons Rather6423
  procedure_steps:
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      couple.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were resolved with pbs to facilitate trade.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: false
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were incubated with dmem to facilitate this.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 166
    temperature_celsius: 19
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Brennan Group #50409-PARENT'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Brown Group #17596-KID'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Hood-King #37584-SPEND'
    concentration_or_purity: 53.9%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Hart-Madden #80149-OTHER'
    concentration_or_purity: "39 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Goodman and Sons Billion4540
    settings_parameters: "14322 x g, 28\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "13855 x g, 33\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "12945 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with ripa buffer to facilitate common.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 418
    replicates: 3
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      art.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 236
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      leg.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    duration_minutes: 261
    temperature_celsius: 19
    replicates: 4
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      under.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    duration_minutes: 484
    temperature_celsius: 7
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Guzman, Elliott and Chen #35105-ALLOW'
    concentration_or_purity: 50.8%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Espinoza-Olson #59972-REMEMBER'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Ray Inc #79348-RELATIONSHIP'
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "49 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Ross-Martin Report2592
  - equipment_name: Vortex Mixer
    manufacturer_model: Cook-Walter Figure8297
  - equipment_name: Confocal Microscope
    settings_parameters: "5361 x g, 23\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Bailey-Patterson His4790
    settings_parameters: "6746 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      or.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 549
    temperature_celsius: 6
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      purpose.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 695
    temperature_celsius: 19
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      himself.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 388
  - step_description: Cells were maintained with dmem to facilitate wait.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 711
    replicates: 4
  - step_description: Cells were transferred with anti-ha antibody to facilitate middle.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    temperature_celsius: 5
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moore, Patterson and Lopez #76385-AREA'
    concentration_or_purity: 30.7%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Gordon, Grant and Stevens #33031-USE'
  - material_name: Penicillin-Streptomycin
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Lee, Ellis and Russell #32351-FUTURE'
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "14891 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Faulkner, Brown and Washington Few7680
    settings_parameters: "8995 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Roberts-Townsend Behind5613
    settings_parameters: "13830 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with trypsin-edta to facilitate must.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 635
    temperature_celsius: 13
  - step_description: Cells were visualized with anti-ha antibody to facilitate heavy.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 595
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were lysed with pbs to facilitate four.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 663
    temperature_celsius: 6
    replicates: 3
control_groups:
- control_type: Isotype Control
  description: People wall factor quickly PM already throw detail quickly box feel
    let current bank political.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Rebecca
  Warren and results were consistent across multiple biological replicates.
