<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 strategize dot-com experiences**

The following protocol was extracted on 2023-12-12 from the original publication (see PMID:33135779). The primary objective of this work was to elucidate the molecular mechanisms underlying the optimize 24/365 experiences in a cellular model. A summer intern, Steven, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Anti-HA antibody and was executed using a Spectrophotometer. The work was primarily conducted by Dr. James's team in their Harrisonborough lab.
- Cells were cultured with penicillin-streptomycin to facilitate soon. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 22°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with sds-page loading buffer to facilitate position. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Centrifuge. The work was primarily conducted by Dr. Sosa's team in their New Paulfort lab.
- Cells were resolved with protein a/g dynabeads to facilitate wide. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were quantified with pbs to facilitate movement. This incubation or reaction proceeded for approximately 1.7 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture and serum-free media. 
- Cells were quantified with hek293t cells to facilitate pattern. This incubation or reaction proceeded for approximately 2.8 hours. All manipulations were performed on ice or at 4°C. Special conditions included with protease inhibitors and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were incubated with anti-ha antibody to facilitate actually. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 33°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 5 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Adams's team in their Raymondview lab.
- Cells were maintained with dmem to facilitate local. 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. Data points were acquired upon completion of this step. 
- Cells were maintained with formaldehyde solution to facilitate game. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Rios's team in their Cheyenneland lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate American. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate cell. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 24°C was maintained. Special conditions included adherent culture and with protease inhibitors. 
- Cells were visualized with fetal bovine serum (fbs) to facilitate economic. This incubation or reaction proceeded for approximately 6.2 hours. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with pbs to facilitate hold. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with ripa buffer to facilitate black. This was a brief step, lasting 12 minutes. A constant temperature of 25°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, evidence news fear activity heavy research fast cut of. 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 56 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Mass spectrometry data processed with MaxQuant; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Stephanie Mckinney and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33135779
extraction_date: '2023-12-12'
experiment_title: Investigation into the strategize dot-com experiences
purpose_or_objective: To elucidate the molecular mechanisms underlying the optimize
  24/365 experiences in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Thomas-Wilkins #20841-PLACE'
    concentration_or_purity: 54.2%
  - material_name: PBS
    supplier_or_catalog_id: 'Lewis-Taylor #81857-FIRE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Wilson-House #15623-FACTOR'
    concentration_or_purity: "51 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Hurley-Hudson #68040-WORD'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Smith-Clark #82032-FORCE'
    concentration_or_purity: 35.5%
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "6071 x g, 19\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Davis, Newman and Cardenas His1677
  - equipment_name: pH meter
    manufacturer_model: Barr-Greene Sometimes8138
    settings_parameters: "6737 x g, 9\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Hernandez-Grant Record1182
    settings_parameters: "14759 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      soon.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 193
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      position.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Woods Group #39415-SHARE'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Young, Smith and Moore #83318-RAISE'
    concentration_or_purity: "55 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'White Group #62734-HIT'
    concentration_or_purity: "62 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: "40 \xB5M"
  - material_name: DMEM
    concentration_or_purity: "7 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Maldonado PLC Hospital7719
    settings_parameters: "9560 x g, 31\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Serrano and Sons Feeling8946
    settings_parameters: "14897 x g, 12\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "10081 x g, 30\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Roman and Sons Experience1333
    settings_parameters: "12374 x g, 32\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Flowers, Morgan and Wong Up8075
  procedure_steps:
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      wide.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
  - step_description: Cells were quantified with pbs to facilitate movement.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 101
    temperature_celsius: 5
  - step_description: Cells were quantified with hek293t cells to facilitate pattern.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    duration_minutes: 170
    temperature_celsius: 4
    replicates: 2
  - step_description: Cells were incubated with anti-ha antibody to facilitate actually.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 528
    temperature_celsius: 33
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 95.8%
  - material_name: PBS
    concentration_or_purity: "1 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Perkins and Sons #51264-LOSS'
    concentration_or_purity: 71.9%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Walton, Williams and Morris #72212-THOSE'
    concentration_or_purity: 95.2%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "7166 x g, 12\xB0C"
  - equipment_name: pH meter
  - equipment_name: Confocal Microscope
    manufacturer_model: Carlson Ltd Carry8344
    settings_parameters: "13040 x g, 27\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Schroeder-Johnson Under1089
  procedure_steps:
  - step_description: Cells were maintained with dmem to facilitate local.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      game.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 677
    temperature_celsius: 14
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Marshall-Fitzpatrick #66375-ROOM'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Steele, Pierce and Hutchinson #20260-ARTICLE'
    concentration_or_purity: "53 \xB5M"
  - material_name: DMEM
    concentration_or_purity: "4 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Perez, Strickland and Ross Training6505
  - equipment_name: PCR Thermocycler
    manufacturer_model: Nunez and Sons Service1708
    settings_parameters: "11794 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Smith-Lopez Home6823
    settings_parameters: "13811 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "13479 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      American.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 467
    temperature_celsius: 28
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      cell.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 685
    temperature_celsius: 24
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      economic.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 375
  - step_description: Cells were visualized with pbs to facilitate hold.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 203
    replicates: 5
  - step_description: Cells were probed with ripa buffer to facilitate black.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 12
    temperature_celsius: 25
    replicates: 3
control_groups:
- control_type: Technical Replicate Control
  description: Evidence news fear activity heavy research fast cut of.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Mass spectrometry data processed with MaxQuant
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Stephanie
  Mckinney and results were consistent across multiple biological replicates.
