Human PPP2R5D knockout A-431 cell lysate (ab270499)
Overview
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Product name
Human PPP2R5D knockout A-431 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; Frameshift = 99.91% -
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 ab280552 - Human PPP2R5D knockout A-431 cell lysate 1 x 100µg ab263973 - Human wild-type A-431 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
Target
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Function
The B regulatory subunit might modulate substrate selectivity and catalytic activity, and also might direct the localization of the catalytic enzyme to a particular subcellular compartment. -
Tissue specificity
Isoform Delta-2 is widely expressed. Isoform Delta-1 is highly expressed in brain. -
Sequence similarities
Belongs to the phosphatase 2A regulatory subunit B56 family. -
Cellular localization
Cytoplasm. Nucleus. Nuclear in interphase, nuclear during mitosis. - Information by UniProt
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Alternative names
- 2A5D_HUMAN
- B'delta
- B56D
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 ab270499 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.
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Notes |
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WB
Use at an assay dependent concentration. |
Images
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Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; Frameshift = 99.91%
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Lane 1: Wild-type A431 cell lysate 20 ug
Lane 2: PPP2R5D knockout A431 cell lysate 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab188325 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188325 was shown to react with PPP2R5D in wild-type A-431 cells in western blot with loss of signal observed in PPP2R5D knockout cell line ab270476 (knockout cell lysate ab270499). Wild-type and PPP2R5D knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188325 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 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. -
Lane 1: Wild-type A431 cell lysate 20 ug
Lane 2: PPP2R5D knockout A431 cell lysate 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab188323 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188323 was shown to react with PPP2R5D in wild-type A-431 cells in western blot with loss of signal observed in PPP2R5D knockout cell line ab270476 (knockout cell lysate ab270499). Wild-type and PPP2R5D knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188323 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 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.
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)
ab270499 has not yet been referenced specifically in any publications.