Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534)

Overview

  • Product name

    Anti-Rab5 antibody [EPR5438] - Early Endosome Marker
    See all Rab5 primary antibodies
  • Description

    Rabbit monoclonal [EPR5438] to Rab5 - Early Endosome Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Rab5 aa 150-250. The exact sequence is proprietary.

  • Positive control

    • 293T, HeLa and Jurkat whole cell lysate (ab7899); Human brain tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109534 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 1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    Images

    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Rab5 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: Hu brain whole cell lysate (20 µg)
      Lane 4: Ms brain whole cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109534 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

    • Immunohistochemical staining of paraffin embedded human pancreas with purified ab109534 at a working dilution of 1 in 1000. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • All lanes : purified at 1/5000 dilution

      Lane 1 : Mouse brain
      Lane 2 : Rat brain

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size: 24 kDa
      Observed band size: 24 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Pancreas tissue sections (with ADM and PanIN lesions) from KC mice labeling Rab5 with ab109534 at 1/200 dilution. Tissue was fixed with paraformaldehyde and permeabilized with TritonX100 0.1% for 10 min at room temperature. Heat-mediated antigen retrieval was performed using TRIS-EDTA pH9 buffer. Tissue sections were blocked with 10% goat serum for 1 hour at 20°C. A polyclonal goat anti-rabbit Alexa Fluor 488 secondary antibody was used at 1/500 dilution. 

      See Abreview

    • All lanes : Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/5000 dilution (purified)

      Lane 1 : MCF7 cell lysate
      Lane 2 : HEK293 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size: 24 kDa
      Observed band size: 24 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/1000 dilution (unpurified)

      Lane 1 : 293T cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : Jurkat cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 24 kDa

    • Immunohistochemical staining of Rab5 in paraffin embedded human brain tissue with unpurified ab109534, at 1/100 dilution.

    References

    This product has been referenced in:

    • Sharoar MG  et al. Sequential formation of different layers of dystrophic neurites in Alzheimer's brains. Mol Psychiatry N/A:N/A (2019). Read more (PubMed: 30899091) »
    • Soung YH  et al. The Role of Arrestin Domain-Containing 3 in Regulating Endocytic Recycling and Extracellular Vesicle Sorting of Integrin ß4 in Breast Cancer. Cancers (Basel) 10:N/A (2018). Read more (PubMed: 30545011) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    1-10 of 10 Abreviews or Q&A

    Application
    Western blot
    Sample
    African green monkey Cell lysate - whole cell (Kidney (COS7 cells))
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Kidney (COS7 cells)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Dec 29 2017

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Dec 29 2017

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Kidney (HEK293 cells))
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Kidney (HEK293 cells)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Dec 29 2017

    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Murine Pancreatic tissue)
    Permeabilization
    Yes - Triton X100 0.01%
    Specification
    Murine Pancreatic tissue
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 25 2017

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Pancreas (with ADM and PanIN lesions) from KC mice)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA pH9
    Permeabilization
    Yes - TritonX100 0.1% 10 min RT
    Specification
    Pancreas (with ADM and PanIN lesions) from KC mice
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 06 2016

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Pancreatic Cancer)
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Loading amount
    30 µg
    Specification
    Pancreatic Cancer
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Aug 01 2016

    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HepaRG)
    Specification
    HepaRG
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.1% TritonX-100
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

    Mrs. Alina Macovei

    Verified customer

    Submitted May 16 2013

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepaRG)
    Loading amount
    20 µg
    Specification
    HepaRG
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

    Mrs. Alina Macovei

    Verified customer

    Submitted May 15 2013

    Answer

    The IgG concentration of Abcam ab109534 lot GR68319 (which is the same as Rab5a 3768-1 Epitomics lot YH112425) is 0.032 mg/ml. This is lower than the IgG concentration of most mouse monoclonal and rabbit polyclonal antibodies but is not unusual for rabbit monoclonals, which generally have much higher affinity for their targets than either mouse monoclonals or rabbit polyclonals.

    Read More

    Answer

    The IgG concentrations of the current lots are:

    3768-1 (ab109534, lot GR68319) - 0.032 mg/ml

    3660-1 (ab125068, lot GR75050) - 1.554 mg/ml

    Read More

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