Human CHI3L1 knockout THP-1 cell lysate (ab280097)
Overview
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Product name
Human CHI3L1 knockout THP-1 cell lysate
See all YKL-40/CHI3L1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
THP-1 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 7 -
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.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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280140 - Human CHI3L1 knockout THP-1 cell lysate 1 x 100µg ab282895 - Human wild-type THP-1 cell lysate 1 x 100µg -
Research areas
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Cell type
acute monocytic leukemia -
Disease
Acute Monocytic Leukemia -
Gender
Male
Target
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Function
Carbohydrate-binding lectin with a preference for chitin. May play a role in defense against pathogens, or in tissue remodeling. May play an important role in the capacity of cells to respond to and cope with changes in their environment. -
Tissue specificity
Present in activated macrophages, articular chondrocytes, synovial cells as well as in liver. Undetectable in muscle tissues, lung, pancreas, mononuclear cells, or fibroblasts. -
Involvement in disease
A genetic variation in CHI3L1 is associated with susceptibility to asthma-related traits type 7 (ASRT7) [MIM:611960]. Asthma-related traits include clinical symptoms of asthma, such as coughing, wheezing and dyspnea, bronchial hyperresponsiveness (BHR) as assessed by methacholine challenge test, serum IgE levels, atopy, and atopic dermatitis. -
Sequence similarities
Belongs to the glycosyl hydrolase 18 family. -
Post-translational
modificationsGlycosylated. -
Cellular localization
Secreted > extracellular space. - Information by UniProt
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Alternative names
- 39 kDa synovial protein
- ASRT7
- Cartilage glycoprotein 39
see all
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280097 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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Western blot - Human CHI3L1 knockout THP-1 cell lysate (ab280097)
Lane 1: Wild-type THP-1 cell lysate 20 μg
Lane 2: CHI3L1 knockout THP-1 cell lysate 20 μg
Lane 3: U-87 MG cell lysate 20 μg
Lane 4: Jurkat cell lysate 20 μg
False colour image of Western blot: Anti-YKL-40/CHI3L1 antibody [EPR19078-157] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab255297 was shown to bind specifically to YKL-40/CHI3L1. A band was observed at 37/40 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CHI3L1 knockout cell line ab280038 (knockout cell lysate ab280097). To generate this image, wild-type and CHI3L1 knockout THP-1 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 CHI3L1 knockout THP-1 cell lysate (ab280097)Lane 1: Wild-type THP-1 cell lysate 20 μg
Lane 2: CHI3L1 knockout THP-1 cell lysate 20 μg
Lane 3: U-87 MG cell lysate 20 μg
Lane 4: Jurkat cell lysate 20 μg
False colour image of Western blot: Anti-YKL-40/CHI3L1 antibody staining at 1 μg/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab77528 was shown to bind specifically to YKL-40/CHI3L1. A band was observed at 33-42 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CHI3L1 knockout cell line ab280038 (knockout cell lysate ab280097). To generate this image, wild-type and CHI3L1 knockout THP-1 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. -
Human CHI3L1 knockout THP-1 cell lysate14 bp deletion in exon 7
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)
ab280097 has not yet been referenced specifically in any publications.