Human DDIT3 knockout SW480 cell lysate (ab270708)
Overview
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Product name
Human DDIT3 knockout SW480 cell lysate
See all DDIT3 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
SW480 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift: 98.84% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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 ab280589 - Human DDIT3 knockout SW480 cell lysate 1 x 100µg ab269601 - Human wild-type SW480 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Male
Target
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Function
Inhibits the DNA-binding activity of C/EBP and LAP by forming heterodimers that cannot bind DNA. -
Involvement in disease
Note=A chromosomal aberration involving DDIT3 is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with FUS. -
Sequence similarities
Belongs to the bZIP family.
Contains 1 bZIP domain. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- C/EBP homologous protein
- C/EBP Homology Protein
- C/EBP zeta
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 ab270708 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 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 19 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 19 kDa. |
Images
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Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift: 98.84%
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Lane 1: Wild-type SW480 cell lysate 20 ug
Lane 2: DDIT3 knockout SW480 cell lysate 20 ug
Lane 3: Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4: HeLa + DMSO control cell lysate 20 ug
Lane 5: HeLa + tunicamycin (20ug/mL,4 hours) cell lysate 20 ug
Lanes 1 - 5: Merged signal (red and green). Green - ab11419 observed at 26 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab11419 was shown to react with DDIT3 in wild-type SW480 cells in western blot with loss of signal observed in DDIT3 knockout cell line ab269585 (knockout cell lysate ab270708). Wild-type and DDIT3 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab11419 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour 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)
ab270708 has not yet been referenced specifically in any publications.