Invitrogen

RAB9 Monoclonal Antibody (Mab9)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

4 Published Figures

19 References

View all (13) RAB9 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

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 Immunocytochemistry (ICC) — Immunofluorescence analysis of RAB9 was performed using HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with RAB9 Mouse Monoclonal Antibody (Product # MA3-067) at 1:200 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in HEK-293 cells. Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of HEK-293 cells showing cytoplasmic (Golgi, ER, endosome pattern) and plasma membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured 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 was performed using Anti-RAB9 Mouse Monoclonal Antibody (Product # MA3-067) and a 23 kDa band corresponding to RAB9 was observed in cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2) and MCF7 (Lane 3) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

RAB9 Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HEK-293 cells were transfected with RAB9A siRNA and reduction of signal was observed in Western Blot using RAB9 Mouse Monoclonal Antibody (Product # MA3-067). {KD}

RAB9 Antibody — Fig. 2 Analysis of endosomal pathway at steady state in fibroblasts. Immunostaining for actin (magenta), lysosomal protein LAMP3 (red), and early endosome protein EEA1 (green) in patient B (Pt. B) and control (PCS Ctl) cells (A i-ii). Immunofluorescence imaging of patient B (Pt. B) and control (PCS Ctl) cells stained for actin (magenta), late endosome maker Rab9 (red), and early endosome protein EEA1 (green) (A iii-iv). Quantification of the average number of early endosomes (p = 8.49E-8) (B), lysosomes (p = 7.09E-28) (C), and late endosomes (not significantly different) (D) in fixed cell images from Pt. B and PCS Ctl cells. DNA counterstaining in blue (Hoechst). Error bars represent SEM. ** p < 0.01 * = p < 0.025. Fig. 2

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1:200-1:500

View 6 publications 6 publications

Immunofluorescence (IF)

1:200-1:500

View 3 publications 3 publications

Immunohistochemistry (IHC)

1:20

View 5 publications 5 publications

Immunomicroscopy (IM)

Assay Dependent

Immunoprecipitation (IP)

Assay dependent

Western Blot (WB)

1:1000-1:5000

View 4 publications 4 publications

Flow Cytometry (Flow)

View 1 publication 1 publication

Product Specifications

Species

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

Published species

Artificial Control, Human, Mouse

Expression System

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

Mab9

Immunogen

Recombinant canine rab 9a.

Conjugate

Unconjugated

Form

Liquid

Concentration

Conc. Not Determined

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; rab9 GTP-binding protein; RAB9, member RAS oncogene family; RAB9-like protein; RAB9A, member RAS oncogene family; Ras-related protein Rab-9A

Gene Aliases: RAB9; RAB9A

UniProt ID: (Human) P51151, (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 Rab protein signal transduction intracellular protein transport metabolic process phagosome-lysosome fusion

Process(es)

GTPase activity protein binding GTP binding GDP binding identical protein binding