Invitrogen

RAB9 Monoclonal Antibody (Mab9)

Product Image Gallery

RAB9 Antibody — Published figure using RAB9 monoclonal antibody (Product # MA3-067)

RAB9 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of RAB9 using RAB9 Monoclonal Antibody (Mab9) (Product # MA3-067) shows staining in A2058 Cells. RAB9 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing RAB9 (Product # MA3-067) at a dilution of 1:100 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

RAB9 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on cancer biopsies of deparaffinized Human prostate carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing RAB9 (Product # MA3-067) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

RAB9 Antibody in Western Blot (WB) — Western blot analysis of RAB9 was performed by loading 25 µg of K562 (lane 1), 293 (lane 2) and Hela (lane 3) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a RAB9 monoclonal antibody (Product # MA3-067) at a dilution of 1:2000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Images were taken at an exposure time of 4 min (lane 1) and 2 min (lanes 2 and 3). Results show a band at ~23 kDa.

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:500

Immunofluorescence (IF)

1:500

Immunohistochemistry (IHC)

1:20

Immunomicroscopy (IM)

Assay Dependent

Immunoprecipitation (IP)

Assay dependent

Western Blot (WB)

1:1000-1:5000

Published Applications

Immunocytochemistry (ICC)

See 4 publications below

Immunofluorescence (IF)

See 1 publication below

Immunohistochemistry (IHC)

See 3 publications below

Flow Cytometry (Flow)

See 1 publication below

Western Blot (WB)

See 3 publications below

Product Specifications

Species Reactivity

Bovine, Dog, Hamster, Human, Non-human primate, Rat

Published species

Human

Host / Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

Mab9

Immunogen

Recombinant canine rab 9a.

Conjugate

Unconjugated

Form

Liquid

Storage buffer

ascites diluted in PBS

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2175599

Product Specific Information

MA3-067 recognizes prenylated and non-prenylated rab 9 from canine, human, monkey, rat, hamster and bovine tissues. This antibody does not cross-react with other rab family members.

MA3-067 has been successfully used in Western blot, immunofluorescence, EM immunocytochemistry and immunoprecipitation procedures. By Western blot, this antibody detects an ~26 kDa protein representing rab 9 from K562 cell extract.

The MA3-067 antigen is recombinant canine rab 9a.

Target Information

Rab 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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: rab-9A; rab9 GTP-binding protein; RAB9, member RAS oncogene family; RAB9A, member RAS oncogene family; Ras-related protein Rab-9; Ras-related protein Rab-9A

Gene Aliases: BOS_26348; RAB9; RAB9A

UniProt ID: (Human) P51151, (Dog) P24408, (Rat) Q99P75

Entrez Gene ID: (Human) 9367, (Bovine) 511776, (Dog) 403947, (Rat) 84589, (Chinese hamster) 100767493

Function(s)

small GTPase mediated signal transduction protein transport regulation of protein localization retrograde transport, endosome to Golgi positive regulation of exocytosis negative regulation by host of symbiont molecular function intracellular protein transport metabolic process Rab protein signal transduction phagosome-lysosome fusion

Process(es)

GTPase activity protein binding GTP binding GDP binding