Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell lysate (ab258080)
Overview
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Product name
Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 5 bp deletion in exon 1. -
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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 ab261135 - Human NPR1 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
Target
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Function
Receptor for the atrial natriuretic peptide NPPA/ANP and the brain natriuretic peptide NPPB/BNP which are potent vasoactive hormones playing a key role in cardiovascular homeostasis. Has guanylate cyclase activity upon binding of the ligand. -
Sequence similarities
Belongs to the adenylyl cyclase class-4/guanylyl cyclase family.
Contains 1 guanylate cyclase domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylation of the protein kinase-like domain is required for full activation by ANP. -
Cellular localization
Membrane. - Information by UniProt
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Alternative names
- ANP-A
- ANPa
- ANPR-A
see all
Associated products
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KO cell lines
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab258080 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. Predicted molecular weight: 119 kDa.
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| Notes |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa. |
Images
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Western blot - Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell lysate (ab258080)
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: NPR1 knockout HeLa cell lysate 20 μg
Lane 3: U-251 MG cell lysate 20 μg
Lane 4: Human Heart tissue lysate 20 μg
Lanes 1 - 4: Merged signal (red and green). Green - anti-NPR1 antibody observed at 140 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
anti-NPR1 antibody was shown to react with NPR1 in wild-type HeLa cells in Western blot with loss of signal observed in NPR1 knockout cell line ab265930 (NPR1 knockout cell lysate ab258080). Wild-type HeLa and NPR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with anti-NPR1 antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 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 h at room temperature before imaging. -
Sanger Sequencing - Human NPR1 knockout HeLa cell lysate (ab258080)Allele-1: 5 bp deletion in exon 1
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Sanger Sequencing - Human NPR1 knockout HeLa cell lysate (ab258080)Allele-2: 1 bp insertion in exon 1
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)
ab258080 has not yet been referenced specifically in any publications.