Human IL13RA2 knockout A375 cell lysate
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IL13RA2 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.
View Alternative Names
CD213a2, Cancer/testis antigen 19, I13R2_HUMAN, IL-13 receptor subunit alpha-2, IL-13R subunit alpha-2, IL-13R-alpha-2, IL-13RA2, IL13 receptor, IL13BP, IL13R, Interleukin 13 receptor alpha 2 chain, Interleukin-13 receptor subunit alpha-2, Interleukin-13-binding protein
- WB
Lab
Western blot - Human IL13RA2 knockout A375 cell lysate (AB275532)
Lane 1 : Wild-type A375 cell lysate 30 ug
Lane 2 : IL13RA2 knockout A375 cell lysate 30 ug
Lane 3 : U-87-MG cell lysate 30 ug
Lane 4 : Daudi cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab260044 observed at 49 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab260044 was shown to react with IL-13 receptor alpha 2 in wild-type A375 cells in western blot with loss of signal observed in IL13RA2 knockout cell line ab273371 (knockout cell lysate ab275532). Wild-type and IL13RA2 knockout A375 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab260044 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (<a href='/en-us/products/primary-antibodies/il-13-receptor-alpha-2-antibody-epr22978-163-ab260044'>ab260044</a>) at 1/1000 dilution
Lane 1:
Wild-type A375 cell lysate at 30 µg
Lane 2:
IL13RA2 knockout A375 cell lysate at 30 µg
Lane 2:
Western blot - Human IL13RA2 knockout A375 cell line (<a href='/en-us/products/cell-lines/human-il13ra2-knockout-a375-cell-line-ab273381'>ab273381</a>)
Lane 3:
U-87-MG cell lysate at 30 µg
Lane 4:
Daudi cell lysate at 30 µg
Predicted band size: 44 kDa
Observed band size: 49 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human IL13RA2 knockout A375 cell lysate (AB275532)
Homozygous : 1 bp deletion in exon 3
- WB
Supplier Data
Western blot - Human IL13RA2 knockout A375 cell lysate (AB275532)
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
Performed under reducing conditions.
In Western blot, ab324598 was shown to bind specifically to IL-13 receptor alpha 2. Band of interest was observed at 50-60 kDa in wild-type A375 cell lysate (lane 1) with no signal observed at this size in IL13RA2 knockout A375 cell lysate (lane 2) (lane 2, knockout cell line ab273381 / knockout cell lysate ab275532).
The molecular weight observed is consistent with what has been described in the literature (PMID : 30718742).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
Exposure time : Lanes 1-2 : 26 seconds, lane 3 : 48 seconds, lane 4 : 180 seconds
All lanes:
Western blot - Anti-IL13RA2 antibody [RM2087] (<a href='/en-us/products/primary-antibodies/il13ra2-antibody-rm2087-ab324598'>ab324598</a>) at 1/1000 dilution
Lane 1:
A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg
Lane 2:
Western blot - Human IL13RA2 knockout A375 cell lysate (ab275532) at 20 µg
Lane 3:
A-172 (human brain glioblastoma cell) whole cell lysate at 20 µg
Lane 4:
Human testis tissue lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 50-60 kDa,36 kDa
false
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB274448
Anti-IL-13 receptor alpha 2 antibody [EPR22978-127] - BSA and Azide free (Capture)
RabMAb|Recombinant
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Product promise
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