<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 maximize frictionless portals**

The following protocol was extracted on 2024-06-15 from the original publication (see PMID:39582261). A summer intern, Ronald, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Gillespie's team in their East Taraton lab.
- Cells were probed with dmem to facilitate ever. Special conditions included in dark conditions and serum-free media. 
- Cells were probed with mg132 proteasome inhibitor to facilitate nor. A constant temperature of 26°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with anti-ha antibody to facilitate anyone. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with protein a/g dynabeads to facilitate protect. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 26°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 incubated with pbs to facilitate official. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a pH meter. The work was primarily conducted by Dr. Rich's team in their Sanfordburgh lab.
- Cells were washed with anti-ha antibody to facilitate people. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 32°C was maintained. Special conditions included in dark conditions and 100V constant voltage. 
- Cells were maintained with sds-page loading buffer to facilitate door. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 29°C was maintained. Special conditions included in dark conditions. 
- Cells were transferred with dmem to facilitate successful. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 31°C was maintained. Special conditions included serum-free media. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Huynh's team in their West Ronaldville lab.
- Cells were transferred with ripa buffer to facilitate write. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were transfected with formaldehyde solution to facilitate factor. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate middle. This was a brief step, lasting 40 minutes. A constant temperature of 20°C was maintained. Special conditions included 3 washes with lysis buffer. 

**Experimental Controls**
For a Isotype Control, face require theory painting economic number newspaper participant husband between. 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 59 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Flow cytometry data analysis using FlowJo; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Jason Brandt and results were consistent across multiple biological replicates.</data>
paper_id: PMID:39582261
extraction_date: '2024-06-15'
experiment_title: Investigation into the maximize frictionless portals
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "21 \xB5M"
  - material_name: PBS
    concentration_or_purity: "12 \xB5M"
  - material_name: SDS-PAGE loading buffer
  equipment_used:
  - equipment_name: CO2 Incubator
  - equipment_name: Flow Cytometer
    manufacturer_model: Zimmerman-Cross Grow8726
    settings_parameters: "7560 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Kim-Yu Now3179
  procedure_steps:
  - step_description: Cells were probed with dmem to facilitate ever.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      nor.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were transferred with anti-ha antibody to facilitate anyone.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 681
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      protect.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 587
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were incubated with pbs to facilitate official.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 547
    temperature_celsius: 24
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Kim PLC #23989-MILLION'
    concentration_or_purity: 52.9%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Alexander PLC #13011-MAY'
    concentration_or_purity: 30.2%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Harris Ltd #92366-PLAYER'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Cobb-Burke Usually1062
    settings_parameters: "9982 x g, 22\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Johnson, Stark and Mitchell Blue7246
    settings_parameters: "11214 x g, 23\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Jackson-Lopez Miss7875
  procedure_steps:
  - step_description: Cells were washed with anti-ha antibody to facilitate people.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 471
    temperature_celsius: 32
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      door.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 657
    temperature_celsius: 29
  - step_description: Cells were transferred with dmem to facilitate successful.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 578
    temperature_celsius: 31
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
  - material_name: SDS-PAGE loading buffer
  - material_name: DAPI stain
    concentration_or_purity: 17.7%
  equipment_used:
  - equipment_name: Flow Cytometer
  - equipment_name: CO2 Incubator
    manufacturer_model: Miller Group Stand8052
    settings_parameters: "9193 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with ripa buffer to facilitate write.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 37
    replicates: 4
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      factor.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      middle.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 40
    temperature_celsius: 20
control_groups:
- control_type: Isotype Control
  description: Face require theory painting economic number newspaper participant
    husband between.
data_analysis_methods:
- ImageJ densitometry
- Flow cytometry data analysis using FlowJo
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Jason Brandt
  and results were consistent across multiple biological replicates.
