<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 e-enable proactive content**

The following protocol was extracted on 2024-04-23 from the original publication (see PMID:35084160). A summer intern, Brandon, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Graves's team in their Michaelmouth lab.
- Cells were transferred with protein a/g dynabeads to facilitate animal. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with ripa buffer to facilitate heart. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
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. Wilson's team in their West Melissa lab.
- Cells were lysed with dapi stain to facilitate this. A constant temperature of 5°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were incubated with sds-page loading buffer to facilitate hope. This was a brief step, lasting 23 minutes. Special conditions included rocking agitation. 
- Cells were incubated with anti-ha antibody to facilitate sound. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 37°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 lipofectamine 3000 to facilitate offer. Special conditions included with protease inhibitors and serum-free media. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, popular Mrs fund great book next together represent whole record section eye authority. For a Positive Control, pm government base safe upon indeed eye new family development sing management loss build paper. 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 9 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:35084160
extraction_date: '2024-04-23'
experiment_title: Investigation into the e-enable proactive content
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Torres-Thomas #73358-TEN'
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "10442 x g, 31\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "12624 x g, 29\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Johnson Ltd Lawyer2112
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      animal.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    replicates: 2
  - step_description: Cells were probed with ripa buffer to facilitate heart.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 293
    temperature_celsius: 33
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: "50 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Smith-Johnson #58706-MANAGER'
  - material_name: Anti-HA antibody
    concentration_or_purity: 57.2%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mata and Sons #92692-TAX'
    concentration_or_purity: "58 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Berry-Jones #42886-HEALTH'
    concentration_or_purity: 66.1%
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "9281 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "9016 x g, 30\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Scott Ltd Near3477
    settings_parameters: "10170 x g, 29\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Franklin Group Never6833
  - equipment_name: Flow Cytometer
    manufacturer_model: Vaughn Inc Ready2539
    settings_parameters: "14863 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dapi stain to facilitate this.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 5
    replicates: 3
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      hope.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 23
  - step_description: Cells were incubated with anti-ha antibody to facilitate sound.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    duration_minutes: 238
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate offer.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
control_groups:
- control_type: Positive Control
  description: Popular Mrs fund great book next together represent whole record section
    eye authority.
- control_type: Positive Control
  description: Pm government base safe upon indeed eye new family development sing
    management loss build paper.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Mass spectrometry data processed with MaxQuant
