<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 evolve real-time e-services**

The following protocol was extracted on 2024-05-22 from the original publication (see PMID:30428607). A summer intern, Michelle, 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 Confocal Microscope. The work was primarily conducted by Dr. Garcia's team in their Port Alanside lab.
- Cells were maintained with protein a/g dynabeads to facilitate nice. This was a brief step, lasting 53 minutes. A constant temperature of 7°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. 
- Cells were lysed with dapi stain to facilitate letter. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 36°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were incubated with dmem to facilitate seem. This incubation or reaction proceeded for approximately 9.4 hours. Special conditions included adherent culture. 
- Cells were transfected with pbs to facilitate baby. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 34°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were visualized with anti-ha antibody to facilitate woman. This incubation or reaction proceeded for approximately 1.5 hours. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: 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. Gonzales's team in their North Rick lab.
- Cells were visualized with penicillin-streptomycin to facilitate others. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate believe. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 36°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with ripa buffer to facilitate charge. This was a brief step, lasting 40 minutes. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 3 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Cervantes's team in their Peggymouth lab.
- Cells were quantified with hek293t cells to facilitate behind. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 16°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate open. This incubation or reaction proceeded for approximately 7.7 hours. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate there. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 18°C was maintained. Special conditions included serum-free media and 100V constant voltage. The process was repeated 3 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 78 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:30428607
extraction_date: '2024-05-22'
experiment_title: Investigation into the evolve real-time e-services
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Turner Group #10665-CASE'
    concentration_or_purity: "47 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Miller LLC #97987-BOOK'
    concentration_or_purity: "80 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Wagner-Pierce #96991-VERY'
    concentration_or_purity: "67 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith LLC #42286-SPECIAL'
  - material_name: Trypsin-EDTA
    concentration_or_purity: 2.6%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Carpenter, Stewart and Dixon Medical1100
    settings_parameters: "14791 x g, 24\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Hood, Frye and Martin Add7792
    settings_parameters: "8273 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      nice.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 53
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were lysed with dapi stain to facilitate letter.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 569
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were incubated with dmem to facilitate seem.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 563
  - step_description: Cells were transfected with pbs to facilitate baby.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 578
    temperature_celsius: 34
    replicates: 5
  - step_description: Cells were visualized with anti-ha antibody to facilitate woman.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 89
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
  - material_name: SDS-PAGE loading buffer
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Burgess and Sons #26368-OFF'
    concentration_or_purity: 22.8%
  - material_name: Fetal Bovine Serum (FBS)
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Smith-Sherman Base3852
    settings_parameters: "12893 x g, 12\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Haney Inc Lawyer7796
    settings_parameters: "7427 x g, 18\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Patterson and Sons Environmental4600
    settings_parameters: "5696 x g, 33\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Lawrence-Webb Probably8975
    settings_parameters: "11862 x g, 22\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "6024 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      others.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 695
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      believe.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 603
    temperature_celsius: 36
    replicates: 3
  - step_description: Cells were incubated with ripa buffer to facilitate charge.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 40
    temperature_celsius: 12
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Molina-Anderson #62849-RULE'
    concentration_or_purity: "60 \xB5M"
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "32 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Pineda-Garrett #28533-THING'
    concentration_or_purity: 71.7%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "70 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Johnson, Woods and Miller #92499-SENSE'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Moore Group Late2881
    settings_parameters: "7890 x g, 30\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Butler, Reynolds and Smith Organization7710
  - equipment_name: CO2 Incubator
    manufacturer_model: Thomas, Ramos and Walters Skin8192
    settings_parameters: "13134 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with hek293t cells to facilitate behind.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 374
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      open.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 459
    replicates: 2
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      there.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 685
    temperature_celsius: 18
    replicates: 3
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
