Human PDCD6IP knockout HEK-293 cell lysate (ab261656)
Overview
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Product name
Human PDCD6IP knockout HEK-293 cell lysate
See all ALIX kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp insertion -
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, 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 ab280406 - Human PDCD6IP knockout HEK293 cell lysate 1 x 100µg ab259780 - Human wild-type HEK-293 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Gender
Female
Target
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Function
Class E VPS protein involved in concentration and sorting of cargo proteins of the multivesicular body (MVB) for incorporation into intralumenal vesicles (ILVs) that are generated by invagination and scission from the limiting membrane of the endosome. Binds to the phospholipid lysobisphosphatidic acid (LBPA) which is abundant in MVBs internal membranes. The MVB pathway appears to require the sequential function of ESCRT-O, -I,-II and -III complexes. The ESCRT machinery also functions in topologically equivalent membrane fission events, such as the terminal stages of cytokinesis and enveloped virus budding (HIV-1 and other lentiviruses). Appears to be an adapter for a subset of ESCRT-III proteins, such as CHMP4, to function at distinct membranes. Required for completion of cytokinesis. Involved in HIV-1 virus budding. Can replace TSG101 it its role of supporting HIV-1 release; this function implies the interaction with CHMP4B. May play a role in the regulation of both apoptosis and cell proliferation. -
Sequence similarities
Contains 1 BRO1 domain. -
Cellular localization
Cytoplasm > cytosol. Melanosome. Cytoplasm > cytoskeleton > centrosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Colocalized with CEP55 in the midbody during cytokinesis. Colocalized with CEP55 at centrosomes of non-dividing cells. - Information by UniProt
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Alternative names
- AIP1
- ALG 2 interacting protein 1
- ALG-2-interacting protein 1
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 ab261656 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.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Western blot - Human PDCD6IP knockout HEK293 cell lysate (ab261656)Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2: ALIX knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab88388 observed at 96 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab88388 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in ALIX knockout cell line ab260864 (knockout cell lysate ab261656). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and ALIX knockout samples were subjected to SDS-PAGE. ab88388 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging. -
Western blot - Human PDCD6IP knockout HEK293 cell lysate (ab261656)Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2: ALIX knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab186728 observed at 96 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab186728 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in ALIX knockout cell line ab260864 (knockout cell lysate ab261656). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and ALIX knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab186728 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 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)
ab261656 has not yet been referenced specifically in any publications.