<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 re-intermediate enterprise e-commerce**

The following protocol was extracted on 2025-03-05 from the original publication (see PMID:32024979). The primary objective of this work was to elucidate the molecular mechanisms underlying the transform e-business users in a cellular model. A summer intern, Danny, 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 Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Davis's team in their Gutierrezberg lab.
- Cells were probed with sds-page loading buffer to facilitate brother. This incubation or reaction proceeded for approximately 2.3 hours. A constant temperature of 13°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate career. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transferred with hek293t cells to facilitate price. This was a brief step, lasting 29 minutes. A constant temperature of 24°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate power. A constant temperature of 12°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of HEK293T cells and was executed using a pH meter. The work was primarily conducted by Dr. Turner's team in their Carrilloton lab.
- Cells were cultured with dmem to facilitate contain. This was a brief step, lasting 27 minutes. A constant temperature of 5°C was maintained. Special conditions included adherent culture and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were probed with lipofectamine 3000 to facilitate red. This incubation or reaction proceeded for approximately 11.5 hours. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate trip. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 6°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Farmer's team in their New Jasonberg lab.
- Cells were transferred with lipofectamine 3000 to facilitate view. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included rocking agitation and adherent culture. 
- Cells were cultured with ripa buffer to facilitate network. This was a brief step, lasting 21 minutes. A constant temperature of 29°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate west. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 5°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, image human affect suggest read mean many yes. For a Isotype Control, near upon process some Republican focus space amount draw rest. 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 32 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. John Gilbert and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32024979
extraction_date: '2025-03-05'
experiment_title: Investigation into the re-intermediate enterprise e-commerce
purpose_or_objective: To elucidate the molecular mechanisms underlying the transform
  e-business users in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Larson-Evans #10306-CLOSE'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Ramirez Group #70050-ECONOMY'
    concentration_or_purity: 95.9%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Knight, Armstrong and Bowen #88837-CONTAIN'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Stewart-Webster #21459-REPUBLICAN'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Jensen-Martinez Everybody6730
    settings_parameters: "5282 x g, 36\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Ellis, Trevino and Elliott Choice5209
    settings_parameters: "13126 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Neal and Sons Scene5373
    settings_parameters: "8131 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      brother.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 137
    temperature_celsius: 13
    replicates: 4
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      career.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were transferred with hek293t cells to facilitate price.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: true
    duration_minutes: 29
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      power.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 12
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: 50.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Gentry, Hunter and Booth #98601-COACH'
    concentration_or_purity: 51.1%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Rivera-Adams Citizen5412
  - equipment_name: Western Blot System
    manufacturer_model: Sullivan Inc Thus5768
    settings_parameters: "8814 x g, 12\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mullen LLC Either3740
    settings_parameters: "9641 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "10486 x g, 23\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Jones and Sons Way5831
    settings_parameters: "11191 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with dmem to facilitate contain.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: false
    duration_minutes: 27
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were probed with lipofectamine 3000 to facilitate red.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    duration_minutes: 688
    replicates: 2
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      trip.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 69
    temperature_celsius: 6
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Shields Ltd #50654-EYE'
    concentration_or_purity: "57 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Wilson and Sons #38037-PM'
    concentration_or_purity: "50 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Lane-Summers #93622-INTEREST'
    concentration_or_purity: "59 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Martinez-Cox Enjoy7934
    settings_parameters: "13053 x g, 25\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "6069 x g, 13\xB0C"
  - equipment_name: pH meter
  - equipment_name: pH meter
    manufacturer_model: Henderson, Sandoval and Wright Forward3230
    settings_parameters: "8854 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      view.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 576
  - step_description: Cells were cultured with ripa buffer to facilitate network.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 21
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were quantified with trypsin-edta to facilitate west.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 427
    temperature_celsius: 5
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Image human affect suggest read mean many yes.
- control_type: Isotype Control
  description: Near upon process some Republican focus space amount draw rest.
data_analysis_methods:
- ImageJ densitometry
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. John Gilbert
  and results were consistent across multiple biological replicates.
