Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJF-R23] to RAB10 - BSA and Azide free
  • Suitable for: WB, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RAB10 antibody [MJF-R23] - BSA and Azide free
    See all RAB10 primary antibodies

  • Description

    Rabbit monoclonal [MJF-R23] to RAB10 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant full length protein corresponding to Human RAB10 aa 1-200. Antibody epitope is C-terminal amino acids DISSGGGVT (AA 185-193)
    Database link: P61026

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • WB: A549, HeLa, HCT 116, MCF7, NIH/3T3, PC-12 and C6 whole cell lysates. ICC/IF: A549 and MCF7 cells. Flow: A549 cells. IP: A549 whole cell lysate.

  • General notes

    Ab238655 is the carrier-free version of ab237703. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab238655 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Western blot - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution

    Lane 1 : Wild-type A549 whole cell lysate
    Lane 2 : RAB10 knockout A549 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 22 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703)

    Lanes 1 - 4: Merged signal (red and green). Green - ab237703 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab237703 was shown to recognize RAB10 in wild-type A549 cells as signal was lost at the expected MW in RAB10 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. Ab237703 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655) Image courtesy of Dr. Dario Alessi from University of Dundee

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue). 

  • Western blot - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Western blot - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution

    Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell line) whole cell lysate
    Lane 2 : Rab10 knockout A549 whole cell lysate
    Lane 3 : HCT 116 (human colorectal carcinoma epithelial cell line) whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
    Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 7 : C6 (rat glial tumor glial 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

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703)

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

    The WT and Rab10 KO A549 lysates were kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in MCF7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) 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 (ab237703).

  • Immunoprecipitation - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Immunoprecipitation - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

    RAB10 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma cell line) whole cell lysate using ab237703 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab237703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution was used for detection.

    Lane 1: A549 whole cell lysate 10μg (input)
    Lane 2: ab237703 IP in A549 whole cell lysate.
    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab237703 in A549 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).

  • Flow Cytometry - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Flow Cytometry - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 with ab237703 at 1/600 dilution (red) compared with the rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody) (blue). Goat Anti-Rabbit IgG H&L (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 (ab237703).

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in A549 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) 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 (ab237703).

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655) Image courtesy of Dr. Dario Alessi from University of Dundee

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized Human A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue). 

  • Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
    Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)

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