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Validated using a knockout cell lineRecombinant

Recombinant Anti-Rab5 antibody [1/Rab5] (ab288769)

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Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
  • Anti-Rab5 antibody [1/Rab5] (ab288769)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [1/Rab5] to Rab5
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Rab5 antibody [1/Rab5]
    See all Rab5 primary antibodies

  • Description

    Mouse monoclonal [1/Rab5] to Rab5

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1, HeLa and MCF7 whole cell lysate. Human brain, heart and kidney lysates. IHC-P: Human astrocytoma. Human breast carcinoma. Human placenta and kidney tissues.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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.

Properties

Associated products

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab288769 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
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 24 kDa.
Notes
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 24 kDa.
Application notes
Is unsuitable for ICC.

Target

  • Function

    Required for the fusion of plasma membranes and early endosomes.

  • Sequence similarities

    Belongs to the small GTPase superfamily. Rab family.

  • Cellular localization

    Cell membrane. Early endosome membrane. Melanosome. Enriched in stage I melanosomes.
  • Information by UniProt

  • Database links

    • Alternative names

      • RAB 5 antibody
      • RAB 5A antibody
      • RAB5A antibody
      • RAB5A member RAS oncogene family antibody
      • RAB5A_HUMAN antibody
      • RAS associated protein RAB5A antibody
      • Ras related protein Rab 5A antibody
      • Ras-related protein Rab-5A antibody
      see all

    Images

    • Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
      All lanes : Anti-Rab5 antibody [1/Rab5] (ab288769) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate
      Lane 2 : Rab5 knockout HAP1 whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution

      Predicted band size: 24 kDa
      Observed band size: 23 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

      Performed under reducing conditions.

      False colour image of Western blot: Anti-Rab5 antibody (ab288769) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab288769 was shown to bind specifically to Rab5. A band was observed at 24 kDa in wild-type HAP1 cell lysates with no signal observed at this size in Rab5 knockout cell lysates. To generate this image, wild-type and Rab5 knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)

      Immunohistochemical analysis of paraffin-embedded human astrocytoma tissue labeling Rab5 with ab288769 at 1/100 (8.35 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human astrocytoma.

      The section was incubated with ab288769 for 30 mins at room temperature and followed by specific mouse IgG2a antibody for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Western blot - Anti-Rab5 antibody [1/Rab5] (ab288769)
      All lanes : Anti-Rab5 antibody [1/Rab5] (ab288769) at 1/1000 dilution

      Lane 1 : Human brain tissue lysate
      Lane 2 : Human heart tissue lysate
      Lane 3 : Human kidney tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

      Predicted band size: 24 kDa
      Observed band size: 23 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

      Exposure time: 70 seconds

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)

      Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Rab5 with ab288769 at 1/100 (8.35 μg/ml) dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human breast carcinoma. The section was incubated with ab288769 for 30 mins at room temperature and followed by specific mouse IgG2a antibody for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The sample was counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Rab5 with ab288769 at 1/100 (8.35 μg/ml) dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human placenta. The section was incubated with ab288769 for 30 mins at room temperature and followed by specific mouse IgG2a antibody for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The sample was counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [1/Rab5] (ab288769)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Rab5 with ab288769 at 1/100 (8.35 μg/ml) dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human kidney. The section was incubated with ab288769 for 30 mins at room temperature and followed by specific mouse IgG2a antibody for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The sample was counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Anti-Rab5 antibody [1/Rab5] (ab288769)
      Anti-Rab5 antibody [1/Rab5] (ab288769)

    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 ab288769? Please let us know so that we can cite the reference in this datasheet.

    ab288769 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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