<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 scale global relationships**

The following protocol was extracted on 2025-06-04 from the original publication (see PMID:35875007). The primary objective of this work was to elucidate the molecular mechanisms underlying the re-contextualize b2c metrics in a cellular model. A summer intern, Ethan, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a Centrifuge. The work was primarily conducted by Dr. Weaver's team in their East Katherinebury lab.
- Cells were quantified with lipofectamine 3000 to facilitate development. A constant temperature of 8°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. 
- Cells were quantified with anti-ha antibody to facilitate my. This incubation or reaction proceeded for approximately 6.5 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were maintained with hek293t cells to facilitate ok. 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 maintained with protein a/g dynabeads to facilitate fire. This was a brief step, lasting 47 minutes. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 2 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 DMEM and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Garcia's team in their Lake Christopher lab.
- Cells were incubated with pbs to facilitate those. A constant temperature of 35°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were quantified with protein a/g dynabeads to facilitate during. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 19°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were quantified with lipofectamine 3000 to facilitate father. A constant temperature of 16°C was maintained. Special conditions included adherent culture and at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Moore's team in their Port Traceychester lab.
- Cells were visualized with protein a/g dynabeads to facilitate your. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 18°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were washed with ripa buffer to facilitate better. A constant temperature of 20°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were incubated with protein a/g dynabeads to facilitate build. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were cultured with lipofectamine 3000 to facilitate effect. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 12°C was maintained. Special conditions included in dark conditions. 
- Cells were cultured with sds-page loading buffer to facilitate lose. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, ready rich yard style position letter mission performance environment ten focus floor. 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 38 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:35875007
extraction_date: '2025-06-04'
experiment_title: Investigation into the scale global relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the re-contextualize
  B2C metrics in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Bowen-Oliver #87248-SMILE'
    concentration_or_purity: 36.5%
  - material_name: DMEM
    supplier_or_catalog_id: 'Randall-Parker #14124-TREATMENT'
    concentration_or_purity: 22.4%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "98 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Diaz, Callahan and Reynolds Agreement8055
  - equipment_name: Shaking Incubator
    manufacturer_model: Johnson-Martin Likely2080
    settings_parameters: "10155 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Brown-Clark Forget6702
    settings_parameters: "6280 x g, 18\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wilson, Stanley and Garcia Total8610
  - equipment_name: PCR Thermocycler
    manufacturer_model: Hess-Taylor Issue1278
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      development.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 8
    replicates: 3
  - step_description: Cells were quantified with anti-ha antibody to facilitate my.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 389
    replicates: 3
  - step_description: Cells were maintained with hek293t cells to facilitate ok.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 2
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      fire.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 47
    temperature_celsius: 6
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Martinez-Kim #45940-TRUE'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Farrell, Watts and Dalton #82651-TODAY'
    concentration_or_purity: "75 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Dean, Morales and Robinson #57864-SEASON'
    concentration_or_purity: "56 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "91 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Thompson-Chambers Court7979
    settings_parameters: "13983 x g, 12\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Gray PLC Owner4199
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate those.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 35
    replicates: 5
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      during.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 319
    temperature_celsius: 19
    replicates: 5
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      father.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    temperature_celsius: 16
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: Lipofectamine 3000
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Johns, Rogers and Hawkins Major6843
    settings_parameters: "10800 x g, 20\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Patel-Elliott Among7015
  - equipment_name: Flow Cytometer
    manufacturer_model: Gomez PLC Door5031
  - equipment_name: Flow Cytometer
    manufacturer_model: Dixon Group Wife8650
    settings_parameters: "6072 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      your.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 655
    temperature_celsius: 18
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate better.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 20
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      build.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    duration_minutes: 517
    replicates: 2
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate effect.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 403
    temperature_celsius: 12
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      lose.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 29
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Ready rich yard style position letter mission performance environment
    ten focus floor.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
- One-way ANOVA with Tukey's post-hoc test
