Human IL1RN (IL1 Receptor Antagonist) knockout A-431 cell lysate (ab275530)
Overview
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Product name
Human IL1RN (IL1 Receptor Antagonist) knockout A-431 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 49 bp deletion in exon 5. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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. -
Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277366 - Human IL1RN knockout A431 cell lysate 1 x 100µg ab277363 - Human wild-type A431 cell lysate 1 x 100µg -
Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
Target
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Relevance
Interleukin 1 receptor antagonist inhibits the activities of interleukin 1 alpha and beta, and modulates a variety of interleukin 1 related immune and inflammatory responses. This gene and five other related cytokine genes form a cluster spanning approximately 400 kb on chromosome 2. A polymorphism of this gene is associated with increased risk of developing osteoporosis and gastric cancer. Four alternatively spliced transcript variants encoding differentt isoforms have been reported. -
Alternative names
- ICIL1RA
- IL1F3
- IL1RA
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275530 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Sanger Sequencing
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Western blot - Human IL1RN knockout A-431 cell lysate (ab275530)Lane 1: Wild-type A431 cell lysate 20 μg
Lane 2: IL-1RA knockout A431 cell lysate 20 μg
False colour image of Western blot: Anti-IL-1RA antibody [EPR6483] staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab124962 was shown to bind specifically to IL-1RA. A band was observed at 19 kDa in wild-type A431 cell lysates with no signal observed at this size in IL1RN knockout cell line ab273379 (knockout cell lysate ab275530). To generate this image, wild-type and IL1RN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween™ 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. -
Western blot - Human IL1RN knockout A-431 cell lysate (ab275530)Lane 1: Wild-type A431 cell lysate 20 μg
Lane 2: IL-1RA knockout A431 cell lysate 20 μg
Lane 3: HeLa cell lysate 20 μg
Lane 4: SK-OV-3 cell lysate 20 μg
Lanes 1 - 4: Merged signal (red and green). Green - Anti-IL-1ra antibody observed at 18 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-IL-1ra antibody was shown to react with IL-1ra in wild-type A431 cells in Western blot with loss of signal observed in IL-1RA knockout cell line ab273379 (IL-1RA knockout cell lysate ab275530). Wild-type A431 and IL-1RA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with anti-IL-1ra antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at 0.5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Donkey anti-Goat IgG H&L (IRDye® 800CW) preabsorbed (ab216775) and Donkey anti-Mouse 680RD secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab275530 has not yet been referenced specifically in any publications.