Human HNRNPA1 knockout HEK-293T cell lysate (ab256942)
Overview
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Product name
Human HNRNPA1 knockout HEK-293T cell lysate
See all hnRNP A1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1. -
Passage number
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.
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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260171 - Human HNRNPA1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Target
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Function
Involved in the packaging of pre-mRNA into hnRNP particles, transport of poly(A) mRNA from the nucleus to the cytoplasm and may modulate splice site selection. May play a role in HCV RNA replication. -
Sequence similarities
Contains 2 RRM (RNA recognition motif) domains. -
Post-translational
modificationsArg-194, Arg-206 and Arg-225 are dimethylated, probably to asymmetric dimethylarginine.
Sumoylated. -
Cellular localization
Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Shuttles continuously between the nucleus and the cytoplasm along with mRNA. Component of ribonucleosomes. In the course of viral infection, colocalizes with HCV NS5B at speckles in the cytoplasm in a HCV-replication dependent manner. - Information by UniProt
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Alternative names
- HNRNPA 1
- Helix destabilizing protein
- Helix-destabilizing protein
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260171 - Human HNRNPA1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Images
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Western blot - Human HNRNPA1 knockout HEK293T cell lysate (ab256942)
Lane 1: Wild-type HEK293T cell lysate (20 µg)
Lane 2: HNRNPA1 knockout HEK293T cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab177152 observed at 37 kDa. Red - loading control ab8245 observed at 36 kDa.
ab177152 Anti-hnRNP A1 antibody [EPR12768] was shown to specifically react with hnRNP A1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266193 (knockout cell lysate ab256942) was used. Wild-type and hnRNP A1 knockout samples were subjected to SDS-PAGE. ab177152 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human HNRNPA1 knockout HEK293T cell lysate (ab256942)Allele-1: 1 bp insertion in exon 1
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Sanger Sequencing - Human HNRNPA1 knockout HEK293T cell lysate (ab256942)Allele-2: 4 bp insertion in exon 1