Overview

  • Product nameAnti-Rab9 antibody [Mab9]
    See all Rab9 primary antibodies
  • Description
    Mouse 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 reactivity
    Reacts with: Mouse, Rat, Hamster, Cow, Dog, Human
    Predicted to work with: Non Human Primates
  • Immunogen

    Recombinant full length protein (Dog).

  • Positive control
    • 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.
  • General notes

    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.

    Alternative versions available:
    Anti-Rab9 antibody (FITC) [Mab9] (ab150011)

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • PurityIgG fraction
  • 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.
  • ClonalityMonoclonal
  • Clone numberMab9
  • IsotypeIgG1
  • Research areas

Applications

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.

Application Abreviews Notes
WB Use a concentration of 10 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
Flow Cyt 1/100.

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.

Target

Anti-Rab9 antibody [Mab9] images

  • 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.

    See Abreview

  • 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.

  • 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.

    Secondary
    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 (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.
  • ab2810 staining Rab9 in MEF by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with PBST and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • 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

References for Anti-Rab9 antibody [Mab9] (ab2810)

This product has been referenced in:
  • Tsai SY  et al. Regulation of TLR3 Activation by S100A9. J Immunol 195:4426-37 (2015). Read more (PubMed: 26385519) »
  • Paul C & Robaire B Impaired function of the blood-testis barrier during aging is preceded by a decline in cell adhesion proteins and GTPases. PLoS One 8:e84354 (2013). WB ; Rat . Read more (PubMed: 24391944) »

See all 13 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (adult testes)
Specification adult testes
Fixative 4% PFA
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Qing Wen

Verified customer

Submitted Mar 27 2013

Thank you for your call today and for letting us know about the trouble with these antibodies.
As we discussed, I've issued credit note *** for the vial of ab51031. The full purchase price will be refunded back to your credit card pending approval...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (pancreatic cancer)
Specification pancreatic cancer
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization Yes - PBS + Triton 0.025%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Dec 09 2010

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (pancreatic cancer)
Specification pancreatic cancer
Fixative Acetone
Permeabilization Yes - PBS + Triton 0.025%
Blocking step MOM kit blocking agaent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Dec 06 2010

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (pancreatic cancer)
Specification pancreatic cancer
Fixative Acetone+Methanol 1:1
Permeabilization Yes - PBST
Blocking step Vector Lab MOM kit #BMK-2202 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Oct 04 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (MEFs)
Specification MEFs
Fixative Formaldehyde
Permeabilization Yes - PBST
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Oct 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (duodenum)
Specification duodenum
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 16 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (pancreatitis)
Loading amount 15 µg
Specification pancreatitis
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (THP-1 cell line)
Loading amount 10 µg
Specification THP-1 cell line
Gel Running Conditions Reduced Denaturing (10% Bis-Tris gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 15 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hela)
Loading amount 100000 cells
Specification Hela
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5 %
Username

Dr. Maria Johansson

Verified customer

Submitted Feb 19 2007

1-10 of 11 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"