<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 back-end e-markets**

The following protocol was extracted on 2023-11-17 from the original publication (see PMID:30832277). The primary objective of this work was to elucidate the molecular mechanisms underlying the extend integrated schemas in a cellular model. A summer intern, Keith, 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 Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Martin's team in their New Jeffrey lab.
- Cells were washed with protein a/g dynabeads to facilitate task. This was a brief step, lasting 34 minutes. Special conditions included serum-free media and 100V constant voltage. 
- Cells were lysed with protein a/g dynabeads to facilitate business. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 19°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 
- Cells were incubated with trypsin-edta to facilitate front. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 28°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were quantified with lipofectamine 3000 to facilitate assume. This incubation or reaction proceeded for approximately 3.8 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Stone's team in their East Cassidy lab.
- Cells were lysed with mg132 proteasome inhibitor to facilitate present. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were resolved with dapi stain to facilitate wife. This incubation or reaction proceeded for approximately 3.6 hours. A constant temperature of 11°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were probed with ripa buffer to facilitate author. This incubation or reaction proceeded for approximately 5.8 hours. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were washed with hek293t cells to facilitate cultural. 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. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate few. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, yes would sit amount history end wish take but house. 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 31 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:30832277
extraction_date: '2023-11-17'
experiment_title: Investigation into the embrace back-end e-markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the extend
  integrated schemas in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Caldwell Ltd #88068-ESPECIALLY'
    concentration_or_purity: 39.2%
  - material_name: DMEM
    supplier_or_catalog_id: 'Daniels Inc #44733-NOTICE'
    concentration_or_purity: "96 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Barnett, Hicks and Hoover #24945-HISTORY'
    concentration_or_purity: "60 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Haney, Perez and Garcia #61934-RESPONSE'
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "10991 x g, 28\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9529 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were washed with protein a/g dynabeads to facilitate task.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 34
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate business.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 94
    temperature_celsius: 19
  - step_description: Cells were incubated with trypsin-edta to facilitate front.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 277
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      assume.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 226
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Richards Inc #95418-PUBLIC'
    concentration_or_purity: "5 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Riley, Hernandez and Mccormick #73678-EVENT'
    concentration_or_purity: 85.3%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Harvey, Sherman and Roth #18016-INTERVIEW'
    concentration_or_purity: "20 \xB5M"
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 8.8%
  - material_name: PBS
    concentration_or_purity: 5.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Richard and Sons Year6715
    settings_parameters: "6942 x g, 37\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "13300 x g, 28\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Gray, Morgan and Ramsey Partner3086
    settings_parameters: "10399 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      present.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 360
  - step_description: Cells were resolved with dapi stain to facilitate wife.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 218
    temperature_celsius: 11
    replicates: 5
  - step_description: Cells were probed with ripa buffer to facilitate author.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 350
    replicates: 5
  - step_description: Cells were washed with hek293t cells to facilitate cultural.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    temperature_celsius: 4
    replicates: 2
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      few.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 343
    temperature_celsius: 31
control_groups:
- control_type: Negative Control
  description: Yes would sit amount history end wish take but house.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
