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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

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Western blot - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
  • Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

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 - BSA and Azide free
  • 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 HRP PE Unconjugated

Overview

  • Product name

    Anti-Rab5 antibody [EPR21801] - BSA and Azide free
    See all Rab5 primary antibodies

  • Description

    Rabbit monoclonal [EPR21801] to Rab5 - BSA and Azide free

  • 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, and MCF7 whole cell lysates.

  • General notes

    ab231095 is the carrier-free version of ab218624.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab231095 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)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.

Target

Images

  • Western blot - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Western blot - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    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.

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

    Developed using the ECL technique.

    Predicted band size: 24 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunoprecipitation - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

  • Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Flow Cytometry (Intracellular) - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624). 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218624).

  • Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)
    Anti-Rab5 antibody [EPR21801] - BSA and Azide free (ab231095)

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 (0)

Publishing research using ab231095? Please let us know so that we can cite the reference in this datasheet.

ab231095 has not yet been referenced specifically in any publications.

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