Human RAB10 knockout A549 cell lysate (ab261677)
Product nameHuman RAB10 knockout A549 cell lysate
See all RAB10 kits
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineA549
Mutation descriptionKnockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
Knockout validationNext Generation Sequencing (NGS), Western Blot (WB)
Reconstitution notesTo 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.
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 applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab280427 - Human RAB10 knockout A549 cell lysate 1 x 100µg ab259782 - Human wild-type A549 cell lysate 1 x 100µg
FunctionThe small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different set of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion (By similarity). That Rab is mainly involved in the biosynthetic transport of proteins from the Golgi to the plasma membrane. Regulates, for instance, SLC2A4/GLUT4 glucose transporter-enriched vesicles delivery to the plasma membrane. In parallel, it regulates the transport of TLR4, a toll-like receptor to the plasma membrane and therefore may be important for innate immune response. Plays also a specific role in asymmetric protein transport to the plasma membrane within the polarized neuron and epithelial cells. In neurons, it is involved in axonogenesis through regulation of vesicular membrane trafficking toward the axonal plasma membrane while in epithelial cells, it regulates transport from the Golgi to the basolateral membrane. Moreover, may play a role in the basolateral recycling pathway and in phagosome maturation. According to PubMed:23263280, may play a role in endoplasmic reticulum dynamics and morphology controlling tubulation along microtubules and tubules fusion.
Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
Cellular localizationCytoplasmic vesicle membrane. Golgi apparatus membrane. Golgi apparatus, trans-Golgi network membrane. Endosome membrane. Recycling endosome membrane. Cytoplasmic vesicle, phagosome membrane. Cell projection, cilium. Endoplasmic reticulum membrane. Associates with SLC2A4/GLUT4 storage vesicles (PubMed:22908308). Localizes to the base of the cilium (PubMed:20576682). Transiently associates with phagosomes (By similarity). Localizes to the endoplasmic reticulum at domains of new tubule growth (PubMed:23263280).
- Information by UniProt
- GTP binding protein RAB10
- RAB10 member RAS oncogene family
KO cell lines
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261677 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2: RAB10 knockout A549 (Human lung carcinoma cell line) whole 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 - ab181367 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab181367 was shown to specifically react with RAB10 in wild-type A549 cells as signal was lost in RAB10 knockout cell line ab261868 (knockout cell lysate ab261677). Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. Ab181367 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.
1 bp insertion
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
ab261677 has not yet been referenced specifically in any publications.