Human RAB29 knockout A549 cell lysate (ab280099)
Overview
-
Product name
Human RAB29 knockout A549 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A549 -
Organism
Human -
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.
-
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 details
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280142 - Human RAB29 knockout A549 cell lysate 1 x 100µg ab277305 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Carcinoma -
Gender
Male
Target
-
Function
Rab GTPase key regulator in vesicle trafficking. Essential for maintaining the integrity of the endosome-trans-Golgi network structure. Together with LRRK2, plays a role in the retrograde trafficking pathway for recycling proteins, such as mannose 6 phosphate receptor (M6PR), between lysosomes and the Golgi apparatus in a retromer-dependent manner. Regulates neuronal process morphology in the intact central nervous system (CNS). May play a role in the formation of typhoid toxin transport intermediates during Salmonella enterica serovar Typhi (S.Typhi) epithelial cell infection. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Post-translational
modificationsIn case of Salmonella enterica serovar Typhimurium (S.Typhimurium) infection, is proteolytically cleaved between Gly-41 and Val-42 by the GtgE viral protease encoded on the Gifsy-2 lysogen bacteriophage, which therefore prevents the recruitment of RAB29 to S.Typhimurium-containing vacuoles. In contrast, no proteolytically cleavage is detected in S.Typhi-infected cells (PubMed:22042847). -
Cellular localization
Cell membrane. Cytoplasm. Cytoplasm, perinuclear region. Golgi apparatus. Golgi apparatus, trans-Golgi network. Vacuole. Cytoplasm, cytoskeleton. Colocalizes with LRRK2 along tubular structures emerging from Golgi apparatus (By similarity). Colocalizes with GM130 at the Golgi apparatus. Colocalizes with dynamic tubules emerging from and retracting to the Golgi apparatus. Colocalizes with TGN46 at the trans-Golgi network (TGN). In Salmonella enterica serovar Typhi (S.Typhi) infected epithelial cells, is recruited and colocalized with both S.Typhi-containing vacuoles and dynamic tubules as well as those emerging from the vacuole toward the cell periphery. - Information by UniProt
-
Alternative names
- DKFZp686P1051
- Rab 7 like protein 1
- RAB 7L
see all
Associated products
-
KO cell lines
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280099 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.
|
Notes |
---|
WB
Use at an assay dependent concentration. |
Images
-
Lane 1: Wild-type A549 lysate 20 μg
Lane 2: RAB29 knockout A549 cell lysate 20 μg
Lane 3: MCF7 cell lysate 20 μg
Lane 4: CACO-2 cell lysate 20 μgFalse colour image of Western blot: Anti-RAB29 antibody [MJF-R30-104] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab256527 was shown to bind specifically to RAB29. A band was observed at 23 kDa in wild-type A549 cell lysates with no signal observed at this size in RAB29 knockout cell line ab280040 (knockout cell lysate ab280099). To generate this image, wild-type and RAB29 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
Lane 1: Wild-type A549 lysate 20 μg
Lane 2: RAB29 knockout A549 cell lysate 20 μg
Lane 3: MCF7 cell lysate 20 μg
Lane 4: CACO-2 cell lysate 20 μgFalse colour image of Western blot: Anti-RAB29 antibody [MJF-R30-124] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab256526 was shown to bind specifically to RAB29. A band was observed at 23 kDa in wild-type A549 cell lysates with no signal observed at this size in RAB29 knockout cell line ab280040 (knockout cell lysate ab280099). To generate this image, wild-type and RAB29 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
Human RAB29 KO in A549 Cells with 119 bp Deletion in Exon 4
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
-
SDS download
-
Datasheet download
References (0)
ab280099 has not yet been referenced specifically in any publications.