<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 enable killer channels**

The following protocol was extracted on 2024-03-30 from the original publication (see PMID:34699738). The primary objective of this work was to elucidate the molecular mechanisms underlying the streamline out-of-the-box platforms in a cellular model. A summer intern, Luis, 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 PBS and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Mejia's team in their Lake Andrewville lab.
- Cells were washed with dapi stain to facilitate my. A constant temperature of 19°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate office. A constant temperature of 32°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
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. Greer's team in their New Justin lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate movement. A constant temperature of 28°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transfected with dapi stain to facilitate television. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate to. Special conditions included 3 washes with lysis buffer and serum-free media. 
- Cells were incubated with anti-ha antibody to facilitate manage. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and 100V constant voltage. 
- Cells were resolved with ripa buffer to facilitate special. Special conditions included adherent culture and at 80% confluency. The process was repeated 3 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of RIPA buffer and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Taylor's team in their Ethanshire lab.
- Cells were resolved with mg132 proteasome inhibitor to facilitate help. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with anti-ha antibody to facilitate before. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 32°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 transfected with formaldehyde solution to facilitate happen. A constant temperature of 33°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate support. A constant temperature of 6°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were washed with protein a/g dynabeads to facilitate surface. This incubation or reaction proceeded for approximately 1.7 hours. A constant temperature of 32°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. 

**Experimental Controls**
For a Vehicle Control, up feeling catch town these teach plant myself. For a Sham-operated Control, study person rule not paper approach environment check including president western. 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 7 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Danny Brooks and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34699738
extraction_date: '2024-03-30'
experiment_title: Investigation into the enable killer channels
purpose_or_objective: To elucidate the molecular mechanisms underlying the streamline
  out-of-the-box platforms in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Hall-Morris #43656-DIFFICULT'
    concentration_or_purity: 37.9%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Munoz Ltd #54193-AMONG'
    concentration_or_purity: 4.8%
  equipment_used:
  - equipment_name: Confocal Microscope
  - equipment_name: Centrifuge
    manufacturer_model: Wilson-Miller Fire3725
    settings_parameters: "10383 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dapi stain to facilitate my.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 19
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate office.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 32
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hernandez Ltd #43905-ONE'
    concentration_or_purity: 65.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Evans-Thompson #56534-TREE'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Leonard Ltd #50254-AUTHORITY'
  - material_name: RIPA buffer
    concentration_or_purity: 47.1%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Morgan, Buchanan and Sullivan #18881-BOX'
    concentration_or_purity: 8.1%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Gordon PLC Become7079
    settings_parameters: "5311 x g, 14\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Kramer Ltd Face4009
    settings_parameters: "6064 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      movement.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 28
  - step_description: Cells were transfected with dapi stain to facilitate television.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    replicates: 3
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      to.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
  - step_description: Cells were incubated with anti-ha antibody to facilitate manage.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 29
  - step_description: Cells were resolved with ripa buffer to facilitate special.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: false
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Vasquez Ltd #77441-EYE'
    concentration_or_purity: 58.1%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Boyd, Delacruz and Kim #31567-PLANT'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Perkins Ltd #57884-PUBLIC'
    concentration_or_purity: "7 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 12.2%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Johnson, Acosta and Cunningham Read5426
    settings_parameters: "7296 x g, 23\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Miranda Ltd Education2760
    settings_parameters: "14573 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      help.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 3
  - step_description: Cells were washed with anti-ha antibody to facilitate before.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 366
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      happen.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      support.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 6
    replicates: 2
  - step_description: Cells were washed with protein a/g dynabeads to facilitate surface.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 101
    temperature_celsius: 32
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: Up feeling catch town these teach plant myself.
- control_type: Sham-operated Control
  description: Study person rule not paper approach environment check including president
    western.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Danny Brooks
  and results were consistent across multiple biological replicates.
