Human RRAGD (Rag D) knockout HEK-293T cell lysate
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RRAGD KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 232 bp deletion in exon1.
View Alternative Names
DKFZp761H171, FLJ44176, OTTHUMP00000016853, RRAGD_HUMAN, Rag D, Rag D protein, RagDRRAGD, Ras related GTP binding D, Ras-related GTP-binding protein D, bA11D8.2.1
- Sanger seq
Unknown
Sanger Sequencing - Human RRAGD (Rag D) knockout HEK-293T cell lysate (AB259091)
Homozygous : 232 bp deletion in exon1
- WB
Lab
Western blot - Human RRAGD (Rag D) knockout HEK-293T cell lysate (AB259091)
Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : RRAGD knockout HEK-293T cell lysate 20 ug
Lane 3 : Jurkat cell lysate 20 ug
Lane 4 : HeLa cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab187679 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab187679 was shown to react with Rag D in wild-type HEK-293T cells in Western blot with loss of signal observed in Rragd knockout cell line ab266287 (Rragd knockout cell lysate ab259091). Wild-type HEK-293T and Rragd knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab187679 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-Rag D antibody - N-terminal (<a href='/en-us/products/primary-antibodies/rag-d-antibody-n-terminal-ab187679'>ab187679</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human RRAGD (Rag D) knockout HEK-293T cell lysate (ab259091) at 20 µg
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 4:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 48 kDa,55 kDa
false
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
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.
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. 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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Rag D participates in the mTORC1 complex a protein complex that senses nutrient availability. It acts as a molecular switch signaling cells to grow or stop growth based on amino acid presence. Rag D facilitates mTORC1 activation by recruiting it to the lysosomal surface where amino acids act as a trigger. This function allows Rag D to play a significant role in cell growth division and metabolism.
Pathways
Rag D integrates into the amino acid sensing pathway directly affecting the mTOR signaling pathway important for cellular growth and homeostasis. Rag D interacts with other Rag proteins such as Rag A and Rag C to form heterodimers that regulate mTORC1 activity. This interaction ensures Rag D can sense and respond to intracellular nutrient levels contributing to homeostatic balance.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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