RAB27A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon5 and 2 bp deletion in exon5 and 5 bp deletion in exon5 and 7 bp insertion in exon5.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon5 and 2 bp deletion in exon5 and 5 bp deletion in exon5 and 7 bp insertion in exon5.
Alternative names
GS2, GTP-binding protein Ram, HsT18676, MGC117246, Mutant Ras related protein Rab-27A, RAB-27A, RAB27A member RAS oncogene family, RAM, RB27A_HUMAN, Rab-27, Ras-related protein Rab-27A
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RAB27A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon5 and 2 bp deletion in exon5 and 5 bp deletion in exon5 and 7 bp insertion in exon5.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon5 and 2 bp deletion in exon5 and 5 bp deletion in exon5 and 7 bp insertion in exon5.
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- RAB27A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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, 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
RAB27A also known as ras-related protein Rab-27A is a small GTPase that plays a role in intracellular vesicle trafficking. It has a molecular mass of approximately 25 kDa. RAB27A localizes in many cell types with higher expressions in secretory cells such as melanocytes and cytotoxic T lymphocytes. It primarily functions in the regulation of vesicle docking and fusion processes with target membranes facilitating the transport of contents such as enzymes and other proteins to specific sites within the cell.
- Biological function summary
This protein regulates the exocytosis processes in certain secretory cells. RAB27A partners with effectors through its GDP-bound and GTP-bound states forming protein complexes to mediate the movement and anchoring of secretory granules. Melanocytes depend on RAB27A to control the distribution of melanosomes which are organelles containing pigment melanin. In immune cells RAB27A regulates the secretion of lytic granules significantly impacting the immune response.
- Pathways
This protein is involved in the melanogenesis pathway and the lytic granule exocytosis pathway. In the melanogenesis pathway RAB27A works closely with proteins such as Melanophilin and Myosin Va to control melanosome transport. In the exocytosis pathway in immune cells RAB27A interacts with proteins like UNC13D which are vital for the release of cytotoxic molecules. By controlling these pathways RAB27A influences pigment formation in skin and hair as well as immune defense mechanisms.
- Associated diseases and disorders
Mutations in the RAB27A gene are associated with Griscelli syndrome type 2 a disorder characterized by partial albinism and immune system defects. This syndrome arises from impaired melanosome transport due to dysfunctional RAB27A and its associated proteins such as Melanophilin and Myosin Va. Additionally defects in RAB27A also relate to hemophagocytic lymphohistiocytosis type 2 where impaired function in cytotoxic T lymphocytes reduces immune response effectiveness leading to excessive inflammation.
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4 product images
Western blot - Human RAB27A knockout A549 cell lysate (ab258618)
Lane 1: Wild-type A549 cell lysate 40 μg
Lane 2: RAB27A knockout A549 cell lysate 40 μg
Lanes 1 - 2: Merged signal (red and green). Green - anti-RAB27A antibody observed at 140 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
anti-RAB27A was shown to react with RAB27A in wild-type A549 cells in Western blot with loss of signal observed in RAB27A knockout cell line Human RAB27A knockout A549 cell line ab266921 (RAB27A knockout cell lysate ab258618). Wild-type A549 and RAB27A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with anti-RAB27A antibody and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.All lanes: anti-RAB27A antibody at 1/1000 dilution
Lanes 1 - 2: RAB27A knockout A549 cell lysate at 40 µg
Lane 2: Western blot - Human RAB27A knockout A549 cell line (Human RAB27A knockout A549 cell line ab266921)
Performed under reducing conditions.
Sanger Sequencing - Human RAB27A knockout A549 cell lysate (ab258618)
Allele-2: 5 bp deletion in exon5
Sanger Sequencing - Human RAB27A knockout A549 cell lysate (ab258618)
Allele-1: 16 bp deletion in exon5
Sanger Sequencing - Human RAB27A knockout A549 cell lysate (ab258618)
Allele-3: 2 bp deletion in exon5
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