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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

Submit a review Submit a question References (7)
Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21801] to Rab5 - Early Endosome Marker
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 APC Carrier Free HRP PE

Overview

  • Product name

    Anti-Rab5 antibody [EPR21801] - Early Endosome Marker
    See all Rab5 primary antibodies

  • Description

    Rabbit monoclonal [EPR21801] to Rab5 - Early Endosome Marker

  • Host species

    Rabbit

  • Specificity

    This antibody could cross-react with RAB5C, but affinity is very low.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, HeLa, MCF7 and Neuro-2a whole cell lysates; Human fetal brain, fetal heart and fetal spleen lysates; Rat and mouse brain and heart lysates. IHC-P: Human kidney and cerebrum tissues; Mouse kidney tissue; Rat liver tissue. ICC/IF: NIH/3T3, HeLa and HAP1 cells. Flow Cyt (intra): MCF7 cells. IP: MCF7 whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab218624 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
Flow Cyt (Intra)
1/50.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/30.
Notes
Flow Cyt (Intra)
1/50.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/30.

Target

Images

  • Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    All lanes : Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624) at 1/1000 dilution

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : Rab5 knockout HAP1 whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 24 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    ab218624 was shown to specifically react with Rab5 in wild-type HAP1 cells as signal was lost in Rab5 knockout cells. Wild-type and Rab5 knockout samples were subjected to SDS-PAGE. Ab218624 and ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/200000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrumentusing the ECL technique.

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilzed wild-type and RAB5A KO HAP1 cells labeling Rab5 with ab218624 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing no staining in RAB5A KO HAP1 cell line and granular cytoplasmic staining in parental HAP1 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Western blot - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    All lanes : Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624) at 1/1000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
    Lane 2 : Human fetal brain lysate
    Lane 3 : Human fetal heart lysate
    Lane 4 : Human fetal spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Mouse brain lysate
    Lane 8 : Mouse heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 24 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?



     

    Exposure time :  Lane 1: 5 seconds; Lanes 2/8: 15 seconds; Lanes 3/4: 6 seconds; Lanes 5/7: 3 seconds;  Lane 6: 26 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Rab5 with ab218624 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in human kidney is observed (PMID 7789520). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Rab5 with ab218624 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in human cerebrum is observed (PMID 7789520). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Rab5 with ab218624 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in mouse kidney is observed (PMID 7789520). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Rab5 with ab218624 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in rat liver is observed (PMID 7789520). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilzed NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Rab5 with ab218624 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing granular cytoplasmic staining in NIH/3T3 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilzed  HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Rab5 with ab218624 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing granular cytoplasmic staining in HeLa cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (human breast adenocarcinoma cell line) cell line labelling Rab5 with ab218624 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

    Rab5 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma cell line) whole cell lysate with ab218624 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218624 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: MCF7 whole cell lysate 10 μg (input).

    Lane 2: ab218624 IP in MCF7 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab218624 in MCF7 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

     

  • Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
    Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (7)

Publishing research using ab218624? Please let us know so that we can cite the reference in this datasheet.

ab218624 has been referenced in 7 publications.

  • Lattao R  et al. Mauve/LYST limits fusion of lysosome-related organelles and promotes centrosomal recruitment of microtubule nucleating proteins. Dev Cell N/A:N/A (2021). PubMed: 33725482
  • Salvany S  et al. Microglial recruitment and mechanisms involved in the disruption of afferent synaptic terminals on spinal cord motor neurons after acute peripheral nerve injury. Glia 69:1216-1240 (2021). PubMed: 33386754
  • Gao Y  et al. Curcumin inhibits classical swine fever virus replication by interfering with lipid metabolism. Vet Microbiol 259:109152 (2021). PubMed: 34146894
  • Sudhakar JN  et al. Lumenal Galectin-9-Lamp2 interaction regulates lysosome and autophagy to prevent pathogenesis in the intestine and pancreas. Nat Commun 11:4286 (2020). PubMed: 32855403
  • Xu DD  et al. Akt+ IKKa/ß+ Rab5+ Signalosome Mediate the Endosomal Recruitment of Sec61 and Contribute to Cross-Presentation in Bone Marrow Precursor Cells. Vaccines (Basel) 8:N/A (2020). PubMed: 32957586
  • Jian Z  et al. Rab5 regulates the proliferation, migration and invasion of glioma cells via cyclin E. Oncol Lett 20:1055-1062 (2020). PubMed: 32724343
  • Wu D & Dean J EXOSC10 sculpts the transcriptome during the growth-to-maturation transition in mouse oocytes. Nucleic Acids Res 48:5349-5365 (2020). PubMed: 32313933

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