Validated using a knockout cell line
Recombinant
RabMAb

Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)

Overview

  • Product name
    Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker
    See all RAB7 primary antibodies
  • Description
    Rabbit monoclonal [EPR7588(B)] to RAB7 - Late Endosome Marker
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human RAB7 aa 50-150. The exact sequence is proprietary.
    Database link: P51149

  • Positive control
    • WB: A375, A431, HACAT, B16F0 and C6 cell lysates and human fetal brain tissue lysate. IHC-P: Human colon adenocarcinoma 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 ab126712 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. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/50 - 1/200.

Target

  • Function
    Key regulator in endo-lysosomal trafficking. Governs early-to-late endosomal maturation, microtubule minus-end as well as plus-end directed endosomal migration and positioning, and endosome-lysosome transport through different protein-protein interaction cascades. Plays a central role, not only in endosomal traffic, but also in many other cellular and physiological events, such as growth-factor-mediated cell signaling, nutrient-transportor mediated nutrient uptake, neurotrophin transport in the axons of neurons and lipid metabolism. Also involved in regulation of some specialized endosomal membrane trafficking, such as maturation of melanosomes, pathogen-induced phagosomes (or vacuoles) and autophagosomes. Plays a role in the maturation and acidification of phagosomes that engulf pathogens, such as S.aureus and M.tuberculosis. Plays a role in the fusion of phagosomes with lysosomes. Plays important roles in microbial pathogen infection and survival, as well as in participating in the life cycle of viruses. Microbial pathogens possess survival strategies governed by RAB7A, sometimes by employing RAB7A function (e.g. Salmonella) and sometimes by excluding RAB7A function (e.g. Mycobacterium). In concert with RAC1, plays a role in regulating the formation of RBs (ruffled borders) in osteoclasts. Controls the endosomal trafficking and neurite outgrowth signaling of NTRK1/TRKA. Regulates the endocytic trafficking of the EGF-EGFR complex by regulating its lysosomal degradation.
  • Tissue specificity
    Widely expressed; high expression found in skeletal muscle.
  • Involvement in disease
    Defects in RAB7A are the cause of Charcot-Marie-Tooth disease type 2B (CMT2B) [MIM:600882]; also known as hereditary motor and sensory neuropathy II (HMSN2). CMT2B is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. CMT2B is clinically characterized by marked distal muscle weakness and a high frequency of foot ulcers, infections and amputations of the toes. CMT2B inheritance is autosomal dominant.
  • Sequence similarities
    Belongs to the small GTPase superfamily. Rab family.
  • Cellular localization
    Late endosome. Lysosome. Cytoplasmic vesicle > phagosome. Melanosome. Cytoplasmic vesicle > phagosome membrane. Co-localizes with OSBPL1A at the late endosome. Found in the ruffled border (a late endosomal-like compartment in the plasma membrane) of bone-resorbing osteoclasts. Recruited to phagosomes containing S.aureus or Mycobacterium.
  • Information by UniProt
  • Database links
  • Alternative names
    • CMT2B antibody
    • PRO2706 antibody
    • PSN antibody
    • RAB7, member RAS oncogene family antibody
    • RAB7A antibody
    • RAB7A, member RAS oncogene family antibody
    • RAB7A_HUMAN antibody
    • Ras associated protein RAB7 antibody
    • Ras related protein Rab7 antibody
    • Ras related protein Rab7a antibody
    • Ras-related protein Rab-7a antibody
    see all

Images

  • ab126712 staining RAB7 in the human cell line A431 (human epidermoid carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)



  • Predicted band size : 23 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: RAB7 knockout HAP1 cell lysate (20 µg)
    Lane 3: A431 cell lysate (20 µg)
    Lane 4: Human fetal brain tissue lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab126712 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab126712 was shown to recognize RAB7 when RAB7 knockout samples were used, along with additional cross-reactive bands. Wild-type and RAB7 knockout samples were subjected to SDS-PAGE. ab126712 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712) at 1/1000 dilution

    Lane 1 : A375 whole cell lysate
    Lane 2 : A431 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
    Blocking and diluting buffer 5% NFDM/TBST
  • All lanes : Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712) at 1/1000 dilution

    Lane 1 : C6 (rat glioma) whole cell lysate
    Lane 2 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryo) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
    Blocking and diluting buffer 5% NFDM/TBST
  • All lanes : Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712) at 1/1000 dilution (unpurified)

    Lane 1 : A375 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : HACAT cell lysate
    Lane 4 : Human fetal brain lysate
    Lane 5 : B16F0 cell lysate
    Lane 6 : C6 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size : 23 kDa
  • Immunohistochemical staining of paraffin-embedded human bladder carcinoma sections labelling RAB7 with purified ab126712 at dilution of 1/70. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunohistochemical analysis of paraffin-embedded Human colon adenocarcinoma tissue labelling Rab7A with unpurified ab126712 at dilution of 1/50.

  • Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HT-29 (human colorectal adenocarcinoma) cells with purified ab126712 at dilution of 1/100. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/1000. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/1000) shown in the top right hand panel. The negative controls are shown in the bottom middle and right hand panels- for negative control 1 rabbit primary antibody and anti-mouse secondary antibody (ab150120) was used. For negative control 2 mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) was used.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Chaubey PM  et al. Proteomic Analysis of the Rat Canalicular Membrane Reveals Expression of a Complex System of P4-ATPases in Liver. PLoS One 11:e0158033 (2016). Read more (PubMed: 27347675) »

See 1 Publication for this product

Customer reviews and Q&As

Cet anticorps n’a pas été testé chez la drosophile et nous n’offrons pas d’échantillon gratuit à des fins de tests préliminaires. Cependant, comme expliqué hier, puisque l'épitop...

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