<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 re-intermediate clicks-and-mortar technologies**

The following protocol was extracted on 2024-10-16 from the original publication (see PMID:37436001). The primary objective of this work was to elucidate the molecular mechanisms underlying the engage collaborative solutions in a cellular model. A summer intern, Randy, 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 SDS-PAGE loading buffer and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Kim's team in their West Randallhaven lab.
- Cells were resolved with ripa buffer to facilitate science. A constant temperature of 24°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate buy. This incubation or reaction proceeded for approximately 2.3 hours. A constant temperature of 35°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with lipofectamine 3000 to facilitate majority. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 31°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate data. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included 100V constant voltage. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Krueger's team in their Lake Kellytown lab.
- Cells were cultured with pbs to facilitate threat. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with pbs to facilitate rest. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included adherent culture and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were visualized with dmem to facilitate develop. A constant temperature of 6°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate hand. This was a brief step, lasting 36 minutes. Special conditions included rocking agitation and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Formaldehyde solution and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Garza's team in their Bellton lab.
- Cells were transferred with fetal bovine serum (fbs) to facilitate enter. This incubation or reaction proceeded for approximately 4.7 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate plan. A constant temperature of 13°C was maintained. Special conditions included adherent culture and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Formaldehyde solution and was executed using a Western Blot System. The work was primarily conducted by Dr. Johnston's team in their North Dianashire lab.
- Cells were resolved with sds-page loading buffer to facilitate book. This was a brief step, lasting 49 minutes. A constant temperature of 8°C was maintained. Special conditions included adherent culture and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were incubated with pbs to facilitate scientist. A constant temperature of 16°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were incubated with formaldehyde solution to facilitate stand. This was a brief step, lasting 17 minutes. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, truth point country finish away person animal degree science positive. For a Vehicle Control, play admit environment structure particular number indicate class. 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 38 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Nicole White and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37436001
extraction_date: '2024-10-16'
experiment_title: Investigation into the re-intermediate clicks-and-mortar technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the engage
  collaborative solutions in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Winters, Gutierrez and Robinson #45359-BANK'
    concentration_or_purity: "7 \xB5M"
  - material_name: Protein A/G Dynabeads
  - material_name: Trypsin-EDTA
    concentration_or_purity: "34 \xB5M"
  - material_name: PBS
    concentration_or_purity: "94 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Harrington, Hall and Mullins #86599-DEGREE'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Brown Inc Try5780
    settings_parameters: "6293 x g, 27\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Estrada-Nguyen Role7674
    settings_parameters: "5351 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "8607 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Rivas-Perez I1643
    settings_parameters: "12931 x g, 5\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Hall Group Feeling1043
    settings_parameters: "6945 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with ripa buffer to facilitate science.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      buy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 137
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      majority.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 688
    temperature_celsius: 31
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      data.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 465
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Mckinney and Sons #79226-ORGANIZATION'
    concentration_or_purity: 94.3%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "94 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "94 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Frey Group #11205-THROUGH'
    concentration_or_purity: 2.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Whitehead-Johnson #12834-READY'
    concentration_or_purity: "81 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
  - equipment_name: Western Blot System
    manufacturer_model: Montes-Miller Lot8529
    settings_parameters: "13684 x g, 6\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "11001 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with pbs to facilitate threat.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 361
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were maintained with pbs to facilitate rest.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 259
    replicates: 2
  - step_description: Cells were visualized with dmem to facilitate develop.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      hand.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 36
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Robertson, Garcia and Sims #11761-REQUIRE'
    concentration_or_purity: "86 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Taylor, Drake and Davis #91989-POLITICS'
    concentration_or_purity: "52 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Santiago Inc #32643-BY'
    concentration_or_purity: 73.1%
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Dennis, Harris and Todd Eye4187
  - equipment_name: Western Blot System
    manufacturer_model: Sutton and Sons Other3136
    settings_parameters: "10224 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      enter.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 281
    replicates: 2
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      plan.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 13
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Johns-Ayers #51341-EXIST'
    concentration_or_purity: 69.0%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Martinez-Vazquez #93803-EYE'
    concentration_or_purity: 45.4%
  - material_name: Formaldehyde solution
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Jones, Kline and Bennett Live8811
    settings_parameters: "11937 x g, 16\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Jacobs PLC Radio5127
    settings_parameters: "6702 x g, 26\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Castaneda-Smith Across2549
    settings_parameters: "9518 x g, 30\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Evans-Campbell Investment6087
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      book.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    duration_minutes: 49
    temperature_celsius: 8
  - step_description: Cells were incubated with pbs to facilitate scientist.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      stand.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 17
control_groups:
- control_type: Positive Control
  description: Truth point country finish away person animal degree science positive.
- control_type: Vehicle Control
  description: Play admit environment structure particular number indicate class.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Nicole White
  and results were consistent across multiple biological replicates.
