Human IPO9 (Importin 9/RANBP9) knockout HeLa cell lysate (ab257483)
Overview
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Product name
Human IPO9 (Importin 9/RANBP9) 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 deletion in exon5 and 26 bp deletion in exon5. -
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 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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262103 - Human IPO9 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
Functions in nuclear protein import as nuclear transport receptor. Serves as receptor for nuclear localization signals (NLS) in cargo substrates. Is thought to mediate docking of the importin/substrate complex to the nuclear pore complex (NPC) through binding to nucleoporin and the complex is subsequently translocated through the pore by an energy requiring, Ran-dependent mechanism. At the nucleoplasmic side of the NPC, Ran binds to the importin, the importin/substrate complex dissociates and importin is re-exported from the nucleus to the cytoplasm where GTP hydrolysis releases Ran. The directionality of nuclear import is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus (By similarity). Mediates the nuclear import of H2B histone (By similarity), RPS7 and RPL18A. Prevents the cytoplasmic aggregation of RPS7 and RPL18A by shielding exposed basic domains. May also import H2A, H3, H4 histones (By similarity), RPL4 and RPL6. -
Sequence similarities
Belongs to the importin beta family.
Contains 1 importin N-terminal domain. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Alternative names
- Imp9
- Imp9a
- Imp9b
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262103 - Human IPO9 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
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
Images
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Western blot - Human IPO9 knockout HeLa cell lysate
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: IPO9 knockout HeLa cell lysate (20 μg)
Lane 3: SK-BR-3 cell lysate (20 μg)
Lanes 1-3: Merged signal (red and green). Green - ab52605. Red - loading control, ab8245 observed at 36 kDa.
ab52605 was shown to react with IPO9 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265352 (knockout cell lysate ab257483) was used. Wild-type and IPO9 knockout samples were subjected to SDS-PAGE. ab52605 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 10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human IPO9 knockout HeLa cell lysateLane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: IPO9 knockout HeLa cell lysate (20 μg)
Lane 3: SK-BR-3 cell lysate (20 μg)
Lanes 1-3: Merged signal (red and green). Green - ab124710. Red - loading control, ab8245 observed at 36 kDa.
ab124710 was shown to react with IPO9 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265352 (knockout cell lysate ab257483) was used. Wild-type and IPO9 knockout samples were subjected to SDS-PAGE. ab124710 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 10000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human IPO9 knockout HeLa cell lysate (ab257483)Allele-1: 26 bp deletion in exon5
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Sanger Sequencing - Human IPO9 knockout HeLa cell lysate (ab257483)Allele-2: 1 bp deletion in exon5
