P01_UoM_CAIX

Project

Project ID
P01_UoM_CAIX

User

Name
Sergi Celis
Institution
University of Manchester

Protein

ID (UniProt code)
Q16790
Source
Human Carbonic Anhydrase IX (38-414), His Tag(CA9-H5226) is expressed from human 293 cells (HEK293). It contains AA Gln 38 - Asp 414
Supplier
Acro Biosystems
Stock (µg)
50.0
State
lyophilised
Protein Buffer PB
20 mM MES, 100 mM NaCl, pH 6.5, trehalose as protectant
Reconstitution olvent
water
Reconstitution volume (µL)
125.0

Selections

  1. Selection P01_UoM_CAIX_SELE_1_1

    Protein

    Input (µg)
    4.8
    Volume (µL)
    12

    Immobilisation

    Beads Source
    Invitrogen_Dynabeads™ His-Tag Isolation and Pulldown_Catalog number 10103D
    Beads (µg)
    160
    Buffer IB
    PB + 0.01% Tween-20
    Incubation (rpm)
    1100
    Temperature (°C)
    4
    Time (minutes)
    30

    Blocking

    Buffer BLB
    IB + 0.2 mg/mL herringsperm DNA
    Volume (µL)
    100

    DEL

    ID(s)
    DEL1-115 spin-filter-purified
    Input (fmol DNA)
    250
    Volume (µL)
    1
    Pre-clearing
    yes
    Incubation (rpm)
    1100
    Temperature (°C)
    23
    Time (minutes)
    45

    Washing

    Buffer
    IB
    Volume (µL)
    100
    Frequency Washes
    10
    Number of washes
    8

    Elution

    Buffer
    water
    Volume (µL)
    20
    Number of elutions
    2
    Incubation (rpm)
    1100
    Temperature (°C)
    80
    Time (minutes)
    5

    PCR

    Buffer
    5 mM Tris/HCl, pH 8.5
    Volume (µL)
    30
    DNA amount (ng)
    600
    Gel image (purified amplicons)
    Selection_1_Gel_Image_Example

    Results

    FASTQ Files - ID
    40-1111632059_P01_UoM_CAIX_SELE_1_1
    FASTQ Files - Reads
    40321
    FASTQ Files - Raw_fastqs
     SELE25-2_R2_001.fastq
    FASTQ Files - Rawdata_qc
     SELE26-2_R2_001.fastq
    BB Enrichment data
     Bck24_R1_001.fastq

Compound Results

  1. Compound Result

    Smiles
    O=S(C1=CC=C(B(O)O)C=C1)(N)=O
    Confirmed Structure
    yes
    On-DNA Validation
     SELE24-1_R1_001.fastq
    Off-DNA Validation
     SELE25-2_R2_001.fastq

  2. Compound Result

    Smiles
    OC[C@@H](O1)[C@@H](O)[C@H](O)[C@@H]2[C@@H]1c3c(O)c(OC)c(O)cc3C(=O)O2
    Confirmed Structure
    no
    On-DNA Validation
     SELE25-2_R2_001.fastq
    Off-DNA Validation
     Bck24_R1_001.fastq