Recombinant
RabMAb

Recombinant Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024)

Overview

  • Product name

    Anti-Annexin-6/ANXA6 antibody [EPR19536]
    See all Annexin-6/ANXA6 primary antibodies
  • Description

    Rabbit monoclonal [EPR19536] to Annexin-6/ANXA6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide within Human Annexin-6/ANXA6 aa 400-500. The exact sequence is proprietary.
    Database link: P08133

    Synthetic peptide within Human Annexin-6/ANXA6 aa 500-600. The exact sequence is proprietary.
    Database link: P08133

  • Positive control

    • WB: Human fetal liver, fetal heart, fetal kidney, spleen and skeletal muscle lysates; Jurkat, Raji, HeLa and HepG2 whole cell lysates. IHC-P: Human tonsil and pancreas tissues. ICC/IF: HepG2 and Raji cells. Flow Cyt: HepG2 cells. IP: Human fetal liver whole cell lysate.
  • General notes

     

     

     This product was previously labelled as Annexin VI

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab201024 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. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa).
ICC/IF 1/500.
IP 1/30.
Flow Cyt 1/600.
IHC-P 1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    May associate with CD21. May regulate the release of Ca(2+) from intracellular stores.
  • Sequence similarities

    Belongs to the annexin family.
    Contains 8 annexin repeats.
  • Domain

    A pair of annexin repeats may form one binding site for calcium and phospholipid.
  • Post-translational
    modifications

    Phosphorylated in response to growth factor stimulation.
  • Cellular localization

    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links

  • Alternative names

    • 67 kDa calelectrin antibody
    • Annexin A6 antibody
    • Annexin VI antibody
    • Annexin VI p68 antibody
    • Annexin-6 antibody
    • AnnexinA6 antibody
    • AnnexinVI antibody
    • ANX 6 antibody
    • ANX A6 antibody
    • ANX6 antibody
    • ANXA 6 antibody
    • ANXA6 antibody
    • ANXA6_HUMAN antibody
    • Calcium binding protein p68 antibody
    • Calelectrin antibody
    • Calphobindin II antibody
    • Calphobindin-II antibody
    • CalphobindinII antibody
    • CBP 68 antibody
    • CBP68 antibody
    • Chromobindin 20 antibody
    • Chromobindin-20 antibody
    • Chromobindin20 antibody
    • CPB II antibody
    • CPB-II antibody
    • CPBII antibody
    • Lipocortin VI antibody
    • LipocortinVI antibody
    • p68 antibody
    • p70 antibody
    • Protein III antibody
    • ProteinIII antibody
    see all

Images

  • All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 76 kDa
    Observed band size: 72 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2 and 3: 5 seconds; Lane 4: 30 seconds.

  • All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024) at 1/5000 dilution

    Lane 1 : Human skeletal muscle lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 5 : HepG2 (Human liver hepatocellular carcinoma 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: 76 kDa
    Observed band size: 72 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2,3,4 and 5: 30 seconds.

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Annexin-6/ANXA6 with ab201024 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on Human tonsil is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

  • Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Annexin-6/ANXA6 with ab201024 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm and membrane staining on the Human pancreas islet is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on HepG2 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab201024 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on Raji cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab201024 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/600 dilution (red) compared with a a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Annexin-6/ANXA6 was immunoprecipitated from 0.35mg of Human fetal liver whole cell lysate with ab201024 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab201024 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Human fetal liver whole cell lysate,10µg (Input).

    Lane 2: ab201024 IP in Human fetal liver whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab201024 in Human fetal liver whole cell lysate.

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

    Exposure time: 3 seconds.

References

ab201024 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Tissue lysate - whole (Soleus muscle)
Gel Running Conditions
Reduced Denaturing (10 % SDS Gel at 15 mA)
Loading amount
7.55 µg
Specification
Soleus muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50µg/mL · Temperature: 23°C

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Verified customer

Submitted Sep 24 2018

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