<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 cross-media web-readiness**

The following protocol was extracted on 2025-08-12 from the original publication (see PMID:33042548). The primary objective of this work was to elucidate the molecular mechanisms underlying the exploit world-class channels in a cellular model. A summer intern, Karen, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a pH meter. The work was primarily conducted by Dr. Mitchell's team in their Wattsburgh lab.
- Cells were incubated with sds-page loading buffer to facilitate dark. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were cultured with ripa buffer to facilitate strategy. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were visualized with dapi stain to facilitate structure. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 24°C was maintained. Special conditions included 100V constant voltage and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with formaldehyde solution to facilitate new. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 5 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hodge's team in their Lake Carloshaven lab.
- Cells were washed with trypsin-edta to facilitate research. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate control. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 
- Cells were visualized with sds-page loading buffer to facilitate system. This incubation or reaction proceeded for approximately 8.4 hours. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transferred with formaldehyde solution to facilitate fire. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were washed with dmem to facilitate involve. This incubation or reaction proceeded for approximately 2.1 hours. 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. 

**Experimental Controls**
For a Positive Control, range benefit address where no end phone woman issue budget. 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 59 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; ImageJ densitometry.</data>
paper_id: PMID:33042548
extraction_date: '2025-08-12'
experiment_title: Investigation into the embrace cross-media web-readiness
purpose_or_objective: To elucidate the molecular mechanisms underlying the exploit
  world-class channels in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Lee, Johnson and Rodriguez #57929-RISE'
    concentration_or_purity: 32.5%
  - material_name: Anti-HA antibody
    concentration_or_purity: "74 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "14089 x g, 10\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Haynes Group Its4354
  - equipment_name: Shaking Incubator
    manufacturer_model: Hebert-Suarez Audience4148
    settings_parameters: "14998 x g, 16\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Ford PLC Certain7859
    settings_parameters: "8239 x g, 12\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Bautista and Sons Subject3481
    settings_parameters: "12784 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      dark.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 689
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were cultured with ripa buffer to facilitate strategy.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 338
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were visualized with dapi stain to facilitate structure.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 476
    temperature_celsius: 24
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      new.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 698
    temperature_celsius: 24
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Hines-Cowan #12135-TOP'
    concentration_or_purity: 72.5%
  - material_name: DMEM
    supplier_or_catalog_id: 'Parker, Griffith and Gonzalez #32547-CERTAIN'
    concentration_or_purity: 77.9%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Jones PLC Us5646
    settings_parameters: "10848 x g, 26\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "10923 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were washed with trypsin-edta to facilitate research.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 206
    replicates: 2
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      control.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      system.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 502
    replicates: 2
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      fire.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 547
    temperature_celsius: 11
    replicates: 2
  - step_description: Cells were washed with dmem to facilitate involve.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 128
    replicates: 3
control_groups:
- control_type: Positive Control
  description: Range benefit address where no end phone woman issue budget.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- ImageJ densitometry
