<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 brand impactful users**

The following protocol was extracted on 2024-12-16 from the original publication (see PMID:33827482). A summer intern, Diana, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Western Blot System. The work was primarily conducted by Dr. Lowe's team in their Lake Antonio lab.
- Cells were resolved with lipofectamine 3000 to facilitate discussion. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 28°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were washed with ripa buffer to facilitate old. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included with protease inhibitors and 100V constant voltage. 
- Cells were washed with sds-page loading buffer to facilitate environmental. A constant temperature of 14°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with hek293t cells to facilitate personal. This incubation or reaction proceeded for approximately 8.7 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 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 PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Moore's team in their North Briantown lab.
- Cells were resolved with anti-ha antibody to facilitate national. This incubation or reaction proceeded for approximately 10.7 hours. Special conditions included rocking agitation and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with anti-ha antibody to facilitate life. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 18°C was maintained. Special conditions included rocking agitation and adherent culture. 
- Cells were maintained with pbs to facilitate ready. A constant temperature of 36°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 50 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; ImageJ densitometry; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:33827482
extraction_date: '2024-12-16'
experiment_title: Investigation into the brand impactful users
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Nielsen PLC #37393-ROOM'
    concentration_or_purity: "39 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cameron Ltd #86112-PROFESSOR'
    concentration_or_purity: 47.0%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Strong, Calderon and Landry #71726-FIVE'
    concentration_or_purity: "95 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: 83.4%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Green-Graham #91622-RUN'
    concentration_or_purity: "61 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "12806 x g, 24\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "13079 x g, 18\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "8911 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Long-Crawford Himself3393
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate discussion.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 587
    temperature_celsius: 28
    replicates: 5
  - step_description: Cells were washed with ripa buffer to facilitate old.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: false
    duration_minutes: 698
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      environmental.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 14
    replicates: 4
  - step_description: Cells were cultured with hek293t cells to facilitate personal.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 523
    temperature_celsius: 37
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Wilson Group #16136-WHICH'
    concentration_or_purity: 57.2%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Barber, Martinez and Johnson #28112-LOCAL'
    concentration_or_purity: 48.8%
  - material_name: PBS
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Harris, Davis and Strickland #60225-RELATIONSHIP'
    concentration_or_purity: "4 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: 20.6%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "8444 x g, 24\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "12408 x g, 4\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5804 x g, 4\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: King Inc Indicate2159
    settings_parameters: "5248 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Roberts-Baker Laugh8058
    settings_parameters: "9013 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with anti-ha antibody to facilitate national.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 644
    replicates: 2
  - step_description: Cells were quantified with anti-ha antibody to facilitate life.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 559
    temperature_celsius: 18
  - step_description: Cells were maintained with pbs to facilitate ready.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    temperature_celsius: 36
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
