Product nameAnti-Rab9 antibody [Mab9]
See all Rab9 primary antibodies
DescriptionMouse monoclonal [Mab9] to Rab9
SpecificityRecognizes prenylated and non-prenylated rab 9. This antibody does not cross-react with other rab family members.
Tested applicationsSuitable for: WB, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Hamster, Cow, Dog, Human
Predicted to work with: Non human primates
Recombinant full length protein (Dog).
- In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; Jurkat; HepG2; K562. This antibody also gave a positive signal in THP1 whole cell lysate (data not shown). In Flow Cytometry, this antibody gave a positive signal in THP1 cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
Primary antibody notesRab proteins are a family of Ras-like GTPases involved in intracellular compartment protein transport. Different members of the 40+ member rab family are responsible for docking and fusion of transport vesicles between different compartments within the cell. Rab 9 is required for trafficking mannose 6-phosphate receptor between the late endosome to trans-Golgi network (TGN). By facilitating receptor transport, rab 9 enables cells to efficiently recycle important cellular trafficking components. It is functionally necessary for rab 9, like other rab family members, to be prenylated by two 20-carbon geranylgeranyl groups at the C-terminus. Most prenylated rab 9 is membrane bound, however, 10-20% of the cellular pool of rab 9 is bound to GDP dissociation inhibitor-alpha (GDI-alpha) in the cytosol. GDI recycles prenylated, GDP bound rab 9 from their fusion targets back to their membranes of origin.
Our Abpromise guarantee covers the use of ab2810 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 10 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).|
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionInvolved in the transport of proteins between the endosomes and the trans Golgi network.
Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
Cellular localizationCell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane.
- Information by UniProt
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All lanes : Anti-Rab9 antibody [Mab9] (ab2810) at 10 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 5 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 22 kDa
Additional bands at: 48 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
ab2810 staining Rab9 in Mouse adult testes tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 4% paraformaldehyde and blocked with 5% BSA for 60 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in 1% BSA) for 15 hours at 4°C. A TRITC-conjugated Donkey anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.
Overlay histogram showing THP1 cells stained with ab2810 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2810, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab2810 (2µg/ml) staining Rab9 in human spleen, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic and cell membrane staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
IHC image of Rab9 staining in human normal tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2810, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
This product has been referenced in:
- Kucera A et al. Spatiotemporal Resolution of Rab9 and CI-MPR Dynamics in the Endocytic Pathway. Traffic 17:211-29 (2016). Read more (PubMed: 26663757) »
- Tsai SY et al. Regulation of TLR3 Activation by S100A9. J Immunol 195:4426-37 (2015). Read more (PubMed: 26385519) »