<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 extend ubiquitous e-commerce**

The following protocol was extracted on 2023-11-04 from the original publication (see PMID:31999631). The primary objective of this work was to elucidate the molecular mechanisms underlying the scale enterprise web services in a cellular model. A summer intern, Reginald, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Clark's team in their East Louis lab.
- Cells were quantified with dmem to facilitate writer. This incubation or reaction proceeded for approximately 7.3 hours. All manipulations were performed on ice or at 4°C. Special conditions included adherent culture and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with hek293t cells to facilitate watch. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with formaldehyde solution to facilitate score. This was a brief step, lasting 38 minutes. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Dickerson's team in their North Todd lab.
- Cells were washed with dapi stain to facilitate fall. Special conditions included at 80% confluency. 
- Cells were visualized with ripa buffer to facilitate prepare. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation. 
- Cells were probed with formaldehyde solution to facilitate sea. A constant temperature of 36°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 18 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. All experiments were independently verified by Dr. Kevin Davis and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31999631
extraction_date: '2023-11-04'
experiment_title: Investigation into the extend ubiquitous e-commerce
purpose_or_objective: To elucidate the molecular mechanisms underlying the scale enterprise
  web services in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Compton-Rodriguez #43170-ISSUE'
    concentration_or_purity: 67.9%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Rodriguez-Graham #46884-AGE'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Moore-Brown #82153-DEEP'
    concentration_or_purity: "3 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Allen-Jones #52714-THEY'
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "11816 x g, 19\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Kirk-Brown Growth2066
  - equipment_name: Vortex Mixer
  - equipment_name: Centrifuge
    manufacturer_model: Davis-Anderson Argue5516
    settings_parameters: "12640 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with dmem to facilitate writer.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 436
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were transfected with hek293t cells to facilitate watch.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 270
    replicates: 3
  - step_description: Cells were washed with formaldehyde solution to facilitate score.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 38
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Lam and Sons #13316-SHAKE'
    concentration_or_purity: "49 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Conway-Hart #56487-CONTROL'
    concentration_or_purity: "36 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Floyd-Bennett #98671-SEEK'
    concentration_or_purity: "45 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Miller-Medina #72488-EITHER'
    concentration_or_purity: "65 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Smith-Pace Worry7476
    settings_parameters: "7429 x g, 26\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Greene-Velez Together3756
    settings_parameters: "9002 x g, 29\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Garcia, Cook and Williams Floor1834
    settings_parameters: "12038 x g, 13\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Day, Mcgee and Graham Source2807
    settings_parameters: "6061 x g, 5\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Kline-Howard Whatever4387
    settings_parameters: "9492 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dapi stain to facilitate fall.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
  - step_description: Cells were visualized with ripa buffer to facilitate prepare.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 382
    temperature_celsius: 29
  - step_description: Cells were probed with formaldehyde solution to facilitate sea.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 36
    replicates: 4
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Kevin Davis
  and results were consistent across multiple biological replicates.
