Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJF-R22-79-3] to RAB8A - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free
    See all RAB8A primary antibodies

  • Description

    Rabbit monoclonal [MJF-R22-79-3] to RAB8A - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

    Recombinant full length protein within Human RAB8A aa 1 to the C-terminus. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.
    Database link: P61006

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

  • Positive control

    • IHC-P: Human bladder cancer and breast tissues. ICC/IF: A549 and HeLa cells. Flow Cyt: A549 cells. IP: A549 whole cell lysate. WB: HeLa and HCT116 cell lysates.

  • General notes

    ab243568 is the carrier-free version of ab241061 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

    Ab243568 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This antibody was developed with support from The Michael J. Fox Foundation.

     

    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-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    All lanes : Anti-RAB8A antibody [MJF-R22-79-3] (ab241061) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RAB8A knockout HeLa cell lysate
    Lane 3 : HCT116 cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab241061).

    Lanes 1-4: Merged signal (red and green). Green - ab241061 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab241061 Anti-RAB8A antibody [MJF-R22-79-3] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab ab241061 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    All lanes : Anti-RAB8A antibody [MJF-R22-79-3] (ab241061) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RAB8A knockout HeLa cell lysate
    Lane 3 : HCT116 cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab241061).

    Lanes 1-4: Merged signal (red and green). Green - ab241061 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab241061 Anti-RAB8A antibody [MJF-R22-79-3] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264994 (knockout cell lysate ab257196) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab241061 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Immunohistochemical analysis of paraffin-embedded human breast tissue labeling RAB8A with ab241061 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human breast (PMID: 14581456). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241061).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling RAB8A with ab241061 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human bladder cancer (PMID: 14581456). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241061).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    RAB8A was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with ab241061 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab241061 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: A549 whole cell lysate 10 µg (Input). 

    Lane 2: ab241061 IP in A549 whole cell lysate . 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab241061 in A549 whole cell lysate .

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 8 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241061).

  • Flow Cytometry - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Flow Cytometry - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cell line labeling RAB8A with ab241061 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary 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 (ab241061).
  • Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RAB8A with ab241061 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody 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 (ab241061).
  • Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB8A with ab241061 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in A549 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody 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 (ab241061).
  • Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

    Immunofluorescent analysis of 3% paraformaldehyde-fixed, 0.1% Saponin permeabilized A549 (human lung carcinoma cell line) and RAB8A knock-out A549 cells labeling RAB8A with ab241061 at 1/100 dilution.

    This image is kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241061).
  • Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
    Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

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