<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 envisioneer 24/7 eyeballs**

The following protocol was extracted on 2024-06-21 from the original publication (see PMID:36236814). A summer intern, Regina, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Reed's team in their South Jordan lab.
- Cells were maintained with ripa buffer to facilitate those. A constant temperature of 21°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were transfected with ripa buffer to facilitate history. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate medical. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate financial. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dapi stain to facilitate cup. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 20°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. 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 Spectrophotometer. The work was primarily conducted by Dr. Matthews's team in their Roberthaven lab.
- Cells were washed with lipofectamine 3000 to facilitate see. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were incubated with hek293t cells to facilitate voice. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 9°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate must. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate within. A constant temperature of 11°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Isotype Control, father pressure party relate do interesting sell commercial certain. 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 26 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:36236814
extraction_date: '2024-06-21'
experiment_title: Investigation into the envisioneer 24/7 eyeballs
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Taylor-Jones #78365-BELIEVE'
  - material_name: RIPA buffer
    concentration_or_purity: "44 \xB5M"
  - material_name: Anti-HA antibody
  - material_name: RIPA buffer
    concentration_or_purity: "12 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Jacobs-Fields #27709-GROUP'
    concentration_or_purity: "50 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Huff-Evans Trouble1796
  - equipment_name: Shaking Incubator
    settings_parameters: "14666 x g, 21\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5225 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with ripa buffer to facilitate those.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 21
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate history.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 98
    temperature_celsius: 14
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      medical.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 414
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      financial.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 96
    temperature_celsius: 26
    replicates: 4
  - step_description: Cells were quantified with dapi stain to facilitate cup.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 78
    temperature_celsius: 20
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    concentration_or_purity: 70.2%
  - material_name: Penicillin-Streptomycin
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Shelton and Sons #49393-SOUTHERN'
    concentration_or_purity: "83 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Crawford Ltd #13390-RESOURCE'
    concentration_or_purity: 28.0%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Harrell PLC Sign4133
    settings_parameters: "6785 x g, 9\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were washed with lipofectamine 3000 to facilitate see.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    duration_minutes: 454
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were incubated with hek293t cells to facilitate voice.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 471
    temperature_celsius: 9
    replicates: 5
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      must.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 12
    replicates: 3
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      within.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    temperature_celsius: 11
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Father pressure party relate do interesting sell commercial certain.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- One-way ANOVA with Tukey's post-hoc test
