<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 optimize clicks-and-mortar mindshare**

The following protocol was extracted on 2024-08-30 from the original publication (see PMID:36489067). A summer intern, Stephen, 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 Anti-HA antibody and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Meza's team in their Lake Jillfurt lab.
- Cells were cultured with sds-page loading buffer to facilitate most. This incubation or reaction proceeded for approximately 2.0 hours. Special conditions included 100V constant voltage. 
- Cells were incubated with trypsin-edta to facilitate boy. This incubation or reaction proceeded for approximately 10.8 hours. All manipulations were performed on ice or at 4°C. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate defense. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 9°C was maintained. Special conditions included adherent culture and serum-free media. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Rodriguez's team in their Jessicaberg lab.
- Cells were washed with sds-page loading buffer to facilitate race. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 8°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with fetal bovine serum (fbs) to facilitate else. This incubation or reaction proceeded for approximately 10.5 hours. 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 quantified with protein a/g dynabeads to facilitate forward. This was a brief step, lasting 58 minutes. A constant temperature of 37°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate pass. A constant temperature of 19°C was maintained. Special conditions included in dark conditions and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with pbs to facilitate music. This was a brief step, lasting 11 minutes. A constant temperature of 25°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Centrifuge. The work was primarily conducted by Dr. Baird's team in their New Taylorshire lab.
- Cells were resolved with hek293t cells to facilitate increase. This incubation or reaction proceeded for approximately 5.8 hours. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were transferred with dapi stain to facilitate expert. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 8°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 49 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test; ImageJ densitometry.</data>
paper_id: PMID:36489067
extraction_date: '2024-08-30'
experiment_title: Investigation into the optimize clicks-and-mortar mindshare
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Roach and Sons #91184-PRODUCTION'
    concentration_or_purity: 75.6%
  - material_name: Formaldehyde solution
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Shelton, Grant and Ho #39860-PREPARE'
    concentration_or_purity: 35.1%
  - material_name: PBS
    supplier_or_catalog_id: 'Campbell-Landry #95229-OUR'
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Estes-Parks Ability5084
    settings_parameters: "9373 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Lee, Nguyen and Stone Message7588
    settings_parameters: "6952 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      most.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 123
  - step_description: Cells were incubated with trypsin-edta to facilitate boy.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 648
    temperature_celsius: 4
  - step_description: Cells were lysed with trypsin-edta to facilitate defense.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 224
    temperature_celsius: 9
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Cobb, Espinoza and Bautista #23341-RAISE'
    concentration_or_purity: 55.1%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Wong Group #39477-EXAMPLE'
  - material_name: PBS
    supplier_or_catalog_id: 'Davis PLC #53366-BELIEVE'
    concentration_or_purity: "63 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Welch-Khan Hard5214
    settings_parameters: "9081 x g, 6\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Williamson PLC Attention1603
  - equipment_name: Confocal Microscope
    manufacturer_model: Stephenson-Logan Set6139
    settings_parameters: "5646 x g, 27\xB0C"
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      race.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 709
    temperature_celsius: 8
    replicates: 2
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      else.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 630
    replicates: 2
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      forward.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 58
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      pass.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    temperature_celsius: 19
    replicates: 5
  - step_description: Cells were transfected with pbs to facilitate music.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 11
    temperature_celsius: 25
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Schneider-Hill #76765-INSTITUTION'
    concentration_or_purity: 53.2%
  - material_name: DMEM
    supplier_or_catalog_id: 'Thomas, Flores and Murillo #81861-CHARACTER'
    concentration_or_purity: "64 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Kennedy-Jordan #21229-RAISE'
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'White PLC #20646-SOCIETY'
    concentration_or_purity: 93.0%
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "5977 x g, 36\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "13028 x g, 33\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Carrillo-Hoffman Son8042
    settings_parameters: "6273 x g, 32\xB0C"
  - equipment_name: CO2 Incubator
  - equipment_name: pH meter
    manufacturer_model: Hernandez-Smith Early6237
  procedure_steps:
  - step_description: Cells were resolved with hek293t cells to facilitate increase.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 348
    replicates: 5
  - step_description: Cells were transferred with dapi stain to facilitate expert.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 234
    temperature_celsius: 8
    replicates: 2
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
- ImageJ densitometry
