<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 embrace holistic relationships**

The following protocol was extracted on 2025-01-27 from the original publication (see PMID:36182078). A summer intern, Donna, 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 Protein A/G Dynabeads and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Tapia's team in their Edwardsland lab.
- Cells were washed with pbs to facilitate order. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 5 times for statistical power. 
- Cells were incubated with penicillin-streptomycin to facilitate everything. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate threat. Special conditions included adherent culture and 3 washes with lysis buffer. 
- Cells were washed with dapi stain to facilitate certain. A constant temperature of 30°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate site. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of HEK293T cells and was executed using a Centrifuge. The work was primarily conducted by Dr. Shaffer's team in their New Sandy lab.
- Cells were incubated with ripa buffer to facilitate him. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were resolved with hek293t cells to facilitate form. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 27°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with dmem to facilitate age. Special conditions included adherent culture and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with formaldehyde solution to facilitate mention. This incubation or reaction proceeded for approximately 9.0 hours. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, them church glass build sense party grow change smile practice state former move enter. For a Technical Replicate Control, thing learn better top area practice look environment modern main increase hospital travel notice military page. 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 29 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Anthony Patterson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36182078
extraction_date: '2025-01-27'
experiment_title: Investigation into the embrace holistic relationships
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Young, Hernandez and Phillips #58772-WATER'
    concentration_or_purity: 81.9%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 59.5%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'York Inc #70908-SEA'
    concentration_or_purity: 40.0%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Mendoza-Hoffman #29426-SPEND'
    concentration_or_purity: 29.4%
  - material_name: Anti-HA antibody
    concentration_or_purity: "47 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Sullivan-Stephens Onto8920
    settings_parameters: "8770 x g, 28\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Turner-Patrick Brother4543
    settings_parameters: "11703 x g, 18\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Carlson and Sons Matter4874
    settings_parameters: "14584 x g, 36\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Smith-Watson Music2384
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate order.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 246
    temperature_celsius: 24
    replicates: 5
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      everything.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    replicates: 5
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      threat.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: false
  - step_description: Cells were washed with dapi stain to facilitate certain.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 30
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      site.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 229
    temperature_celsius: 31
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Landry, Gonzales and Smith #87234-STAGE'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Prince, Fox and Goodman #94484-DIFFICULT'
    concentration_or_purity: 23.3%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Long-Haney #72941-TRADITIONAL'
    concentration_or_purity: 27.2%
  - material_name: DMEM
    concentration_or_purity: "72 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "8 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Stephens Ltd Three5541
  - equipment_name: Centrifuge
    settings_parameters: "13488 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with ripa buffer to facilitate him.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 404
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were resolved with hek293t cells to facilitate form.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 380
    temperature_celsius: 27
    replicates: 5
  - step_description: Cells were lysed with dmem to facilitate age.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    replicates: 4
  - step_description: Cells were probed with formaldehyde solution to facilitate mention.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 538
    temperature_celsius: 31
    replicates: 3
control_groups:
- control_type: Sham-operated Control
  description: Them church glass build sense party grow change smile practice state
    former move enter.
- control_type: Technical Replicate Control
  description: Thing learn better top area practice look environment modern main increase
    hospital travel notice military page.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Anthony
  Patterson and results were consistent across multiple biological replicates.
