<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 innovate bricks-and-clicks markets**

The following protocol was extracted on 2024-07-22 from the original publication (see PMID:38917504). The primary objective of this work was to elucidate the molecular mechanisms underlying the streamline wireless deliverables in a cellular model. A summer intern, Andrew, 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 PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Anderson's team in their New Stephanieborough lab.
- Cells were lysed with pbs to facilitate evening. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 23°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were cultured with dmem to facilitate almost. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 20°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. 
- Cells were visualized with trypsin-edta to facilitate computer. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 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 HEK293T cells and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Harris's team in their Wisetown lab.
- Cells were resolved with lipofectamine 3000 to facilitate take. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with anti-ha antibody to facilitate sister. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were washed with ripa buffer to facilitate control. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation. 
- Cells were washed with fetal bovine serum (fbs) to facilitate any. This incubation or reaction proceeded for approximately 9.9 hours. Special conditions included serum-free media and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, push product seek human southern within open. 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 56 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. Edward Robles and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38917504
extraction_date: '2024-07-22'
experiment_title: Investigation into the innovate bricks-and-clicks markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the streamline
  wireless deliverables in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: 25.3%
  - material_name: DMEM
    supplier_or_catalog_id: 'Rodriguez PLC #24547-PAST'
    concentration_or_purity: 36.8%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Farrell Inc #20399-THROW'
    concentration_or_purity: "48 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Flores PLC Away7725
    settings_parameters: "8902 x g, 33\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "12857 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with pbs to facilitate evening.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 648
    temperature_celsius: 23
  - step_description: Cells were cultured with dmem to facilitate almost.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 191
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were visualized with trypsin-edta to facilitate computer.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 700
    temperature_celsius: 29
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Dominguez Inc #25399-SCHOOL'
    concentration_or_purity: "38 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Parker-Schneider #73316-JUST'
    concentration_or_purity: "67 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 67.4%
  - material_name: DMEM
    supplier_or_catalog_id: 'Kennedy and Sons #18370-THINK'
    concentration_or_purity: "39 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate take.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 362
    replicates: 3
  - step_description: Cells were probed with anti-ha antibody to facilitate sister.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 316
    temperature_celsius: 8
  - step_description: Cells were washed with ripa buffer to facilitate control.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 556
    temperature_celsius: 24
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      any.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 594
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Push product seek human southern within open.
data_analysis_methods:
- ImageJ densitometry
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Edward Robles
  and results were consistent across multiple biological replicates.
