Validated using a knockout cell line
Recombinant
RabMAb

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

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7588(B)] to RAB7 - Late Endosome Marker
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide 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. ICC/IF: HT-29 cells.

  • 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.

    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.

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 IHC antigen retrieval protocols.
Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/50 - 1/500.

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

    see all

  • 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

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

    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.

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

    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)

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)

     Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma epithelial cell) labeling RAB7 with ab126712 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. 

    Confocal image showing cytoplasmic staining in HT-29 cells.

     

  • Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    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
    All lanes : 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
  • Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    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
    All lanes : 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
  • Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    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
    All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 23 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)

    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.

  • Other - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Other - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (ab126712)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References (1)

Publishing research using ab126712? Please let us know so that we can cite the reference in this datasheet.

ab126712 has been referenced in 1 publication.

  • 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). PubMed: 27347675

Customer reviews and Q&As

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Question

Suite à mon appel au support scientifique cet après midi, je souhaiterai tester l'anticorps ab 126712 (anti-Rab7) humain chez l'espèce drosophile. En effet, l'épitope reconnu par cet anticorps correspond à une région avec plus de 85% d'homologie chez la drosophile. Serait-il possible de m'envoyer un échantillon gratuit de cet anticorps, comme vous me l'avez proposé au téléphone ?

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Answer

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'épitope reconnu par cet anticorps correspond à une région avec plus de 85% d'homologie chez la drosophile, je peux vous faire une remise si vous souhaitez tester l’anticorps.

Je suis donc ravie que vous souhaitiez participer à notre offre et tester ab126712 chez la drosophile. Comme expliqué au téléphone, après soumission d’une Abreview avec vos résultats, un avoir de la valeur de l'anticorps testé vous sera offert.

Code de remise : xxxxxxx
Date d’expiration : 07/04/13

Le code de réduction correspondant est xxxxxxxxx. Ce code de réduction est utilisable sous 120 jours et pour l’achat d’un autre produit.

Vous devez dans un premier temps acheter l'anticorps puis soumettre une Abreview avec vos résultats chez la drosophile en notant ce code dans la section « Additional notes ». Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de ab126712.

Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.

Termes et Conditions : www.abcam.com/collaborationdiscount.

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