Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Rab5b antibody (ab72907)

Key features and details

  • Rabbit polyclonal to Rab5b
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Rab5b antibody
    See all Rab5b primary antibodies

  • Description

    Rabbit polyclonal to Rab5b

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Orangutan

  • Immunogen

    Synthetic peptide corresponding to Human Rab5b aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab102726)

  • Positive control

    • This antibody gave a positive signal in the following lysates: Human Placenta Tissue; HeLa Whole Cell; Jurkat Whole Cell; HEK293 Whole Cell; HepG2 Whole Cell; Human Brain Tissue.

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab72907 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 mg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).

Target

  • Relevance

    Rab5b is a member of the Rab family of small (monomeric) G proteins. Like other small G proteins, Rab5b switches between an inactive, GDP-form and an active, GTP-bound form. GDP/GTP exchange factors (GEFs) catalyse the conversion from the GDP-bound form to the GTP-bound form, while GTPase-activating proteins (GAPs) catalyse GTP hydrolysis to GDP. Rab5b is involved in endocytosis and recycling of cell surface molecules. It interacts with RIN2 and RIN3, which regulate its function, possibly by acting as GEFs. Knockdown of Rab5b abolished group I metabotropic glutamate receptor (mGluR)-mediated neuroprotection. Furthermore, Rab5b interacts with LRRK2, the defective gene at the PARK8 locus that results in Parkinson's disease. Roles for Rab5b in neurodegenerative disease, neuroprotection, and synaptic plasticity have been suggested.

  • Cellular localization

    Cell Membrane, Endosome

  • Database links

    see all

  • Alternative names

    • Rab 5b antibody
    • Rab5 b antibody
    • RAB5B, member RAS oncogene family antibody
    • Ras related protein Rab 5B antibody
    • RAS-associated protein RAB5B antibody
    see all

Images

  • Western blot - Anti-Rab5b antibody (ab72907)
    Western blot - Anti-Rab5b antibody (ab72907)
    All lanes : Anti-Rab5b antibody (ab72907) at 1 µg/ml

    Lane 1 : Human placenta tissue lysate - total protein (ab29745)
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa, 70 kDa, 90 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes


    Rab5b contains a number of potential lipidation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5b antibody (ab72907)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5b antibody (ab72907)
    ICC/IF image of ab72907 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72907, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.

References (1)

Publishing research using ab72907? Please let us know so that we can cite the reference in this datasheet.

ab72907 has been referenced in 1 publication.

  • Connor-Robson N  et al. An integrated transcriptomics and proteomics analysis reveals functional endocytic dysregulation caused by mutations in LRRK2. Neurobiol Dis 127:512-526 (2019). PubMed: 30954703

Images

  • Western blot - Anti-Rab5b antibody (ab72907)
    Western blot - Anti-Rab5b antibody (ab72907)
    All lanes : Anti-Rab5b antibody (ab72907) at 1 µg/ml

    Lane 1 : Human placenta tissue lysate - total protein (ab29745)
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa, 70 kDa, 90 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes


    Rab5b contains a number of potential lipidation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5b antibody (ab72907)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5b antibody (ab72907)
    ICC/IF image of ab72907 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72907, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.

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