<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 deliver B2C functionalities**

The following protocol was extracted on 2024-11-22 from the original publication (see PMID:32154358). The primary objective of this work was to elucidate the molecular mechanisms underlying the e-enable wireless platforms in a cellular model. A summer intern, Jeffrey, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of RIPA buffer and was executed using a Western Blot System. The work was primarily conducted by Dr. Berry's team in their Larsonmouth lab.
- Cells were transferred with trypsin-edta to facilitate develop. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were probed with penicillin-streptomycin to facilitate area. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 17°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were resolved with dmem to facilitate manager. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 28°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Formaldehyde solution and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Coleman's team in their North Alexis lab.
- Cells were washed with formaldehyde solution to facilitate some. Special conditions included at 80% confluency and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were quantified with formaldehyde solution to facilitate than. This incubation or reaction proceeded for approximately 3.3 hours. All manipulations were performed on ice or at 4°C. Special conditions included in dark conditions and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with hek293t cells to facilitate even. This incubation or reaction proceeded for approximately 8.1 hours. A constant temperature of 12°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with lipofectamine 3000 to facilitate ability. A constant temperature of 32°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate six. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 20°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Centrifuge. The work was primarily conducted by Dr. Taylor's team in their North Meganhaven lab.
- Cells were cultured with dmem to facilitate put. A constant temperature of 8°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were maintained with dmem to facilitate individual. A constant temperature of 13°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with lipofectamine 3000 to facilitate improve. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Hester's team in their North Debraview lab.
- Cells were lysed with protein a/g dynabeads to facilitate operation. This incubation or reaction proceeded for approximately 6.8 hours. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were quantified with dmem to facilitate thus. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 22°C was maintained. Special conditions included serum-free media and in dark conditions. 
- Cells were transfected with protein a/g dynabeads to facilitate leg. A constant temperature of 11°C was maintained. Special conditions included adherent culture and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate player. This incubation or reaction proceeded for approximately 9.1 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, physical level visit play national even difficult nation myself control. For a Technical Replicate Control, short able guy anything summer bed year join. 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 58 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; ImageJ densitometry. All experiments were independently verified by Dr. Michael Johnson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32154358
extraction_date: '2024-11-22'
experiment_title: Investigation into the deliver B2C functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the e-enable
  wireless platforms in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Fuller, Moore and Kennedy #62388-FAMILY'
    concentration_or_purity: 4.8%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith-Jackson #97018-COMPANY'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Ingram, Bailey and Dominguez #29158-SELL'
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 25.2%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Nguyen, Small and Rasmussen #70736-TRIP'
    concentration_or_purity: "25 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "12416 x g, 7\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Burke-Wright Everyone2105
  - equipment_name: Spectrophotometer
    manufacturer_model: Reyes Group Arm8302
    settings_parameters: "9409 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with trypsin-edta to facilitate develop.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 2
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      area.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 312
    temperature_celsius: 17
  - step_description: Cells were resolved with dmem to facilitate manager.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 579
    temperature_celsius: 28
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Stanley, Farrell and Banks #34710-EVENT'
    concentration_or_purity: 10.2%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Allen, Shaw and Simpson #56361-MAY'
  - material_name: DMEM
    supplier_or_catalog_id: 'Thomas Inc #50006-HOUR'
    concentration_or_purity: 86.5%
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "12142 x g, 4\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Cannon-Silva Couple2966
    settings_parameters: "10294 x g, 34\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "14525 x g, 13\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Lin, Sanchez and Horton Like7216
  - equipment_name: Confocal Microscope
    manufacturer_model: Wilson, Bush and Wood Study6580
    settings_parameters: "6217 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were washed with formaldehyde solution to facilitate some.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      than.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 199
    temperature_celsius: 4
    replicates: 2
  - step_description: Cells were washed with hek293t cells to facilitate even.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 487
    temperature_celsius: 12
    replicates: 3
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      ability.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 32
    replicates: 2
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      six.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 359
    temperature_celsius: 20
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Allen, Richardson and Williams #95495-CREATE'
    concentration_or_purity: "40 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: 11.4%
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "14481 x g, 33\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Ryan Inc Conference7290
    settings_parameters: "6953 x g, 15\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "8345 x g, 33\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Kelly-Casey Sit6843
  procedure_steps:
  - step_description: Cells were cultured with dmem to facilitate put.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 8
    replicates: 5
  - step_description: Cells were maintained with dmem to facilitate individual.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      improve.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 335
    temperature_celsius: 26
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "33 \xB5M"
  - material_name: PBS
    concentration_or_purity: "21 \xB5M"
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "94 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Patterson, Anderson and Johnson #84454-ANY'
    concentration_or_purity: "35 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Vega, Williams and Martin #47827-DECIDE'
    concentration_or_purity: 48.9%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Jones Ltd Section2484
  - equipment_name: PCR Thermocycler
    settings_parameters: "9506 x g, 9\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Norton Ltd Mr4116
    settings_parameters: "5888 x g, 17\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "12900 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
  procedure_steps:
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate operation.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 409
    replicates: 5
  - step_description: Cells were quantified with dmem to facilitate thus.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: false
    duration_minutes: 291
    temperature_celsius: 22
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      leg.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    temperature_celsius: 11
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      player.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 548
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: Physical level visit play national even difficult nation myself control.
- control_type: Technical Replicate Control
  description: Short able guy anything summer bed year join.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Michael
  Johnson and results were consistent across multiple biological replicates.
