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  1. Link

    annexin-6anxa6-antibody-epr19517-ab201023.pdf

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Signal Transduction Cytoskeleton / ECM Cell Adhesion Annexins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Immunoprecipitation - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
  • Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19517] to Annexin-6/ANXA6
  • Suitable for: IHC-P, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Annexin-6/ANXA6 antibody [EPR19517]
    See all Annexin-6/ANXA6 primary antibodies
  • Description

    Rabbit monoclonal [EPR19517] to Annexin-6/ANXA6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal liver, fetal heart, fetal kidney and fetal spleen lysates; A549, HeLa, Jurkat, Raji, C6, NIH/3T3 and PC-12 whole cell lysates; Mouse brain, heart, spleen and kidney lysates; Rat brain, heart, kidney and spleen lysates. IHC-P: Human tonsil, pancreas and stomach tissues. IP: Human fetal liver whole cell lysate.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19517
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Annexins
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calcium Binding Proteins

Associated products

  • Alternative Versions

    • Anti-Annexin-6/ANXA6 antibody [EPR19517] - BSA and Azide free (ab240359)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ANXA6 (Annexin-6) knockout HeLa cell line (ab265677)
  • KO cell lysates

    • Human ANXA6 (Annexin-6) knockout HeLa cell lysate (ab257351)
  • Recombinant Protein

    • Recombinant Human Annexin-6/ANXA6 protein (ab92968)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201023 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
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa).
IP
1/40.
Notes
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa).
IP
1/40.

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.
  • Target information above from: UniProt accession P08133 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 309 Human
    • Entrez Gene: 11749 Mouse
    • Entrez Gene: 79125 Rat
    • Omim: 114070 Human
    • SwissProt: P08133 Human
    • SwissProt: P14824 Mouse
    • SwissProt: P48037 Rat
    • Unigene: 412117 Human
    • Unigene: 265347 Mouse
    • Unigene: 204053 Rat
    see all
  • 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

  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ANXA6 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Raji cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 76 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab201023 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab201023 Anti-Annexin-6/ANXA6 antibody [EPR19536] was shown to specifically react with 6/ANXA6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265677 (knockout cell lysate ab257351) was used.  Wild-type and 6/ANXA6 knockout samples were subjected to SDS-PAGE.  ab201023 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on lymphocytes of 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

    Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Membrane staining on 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

    Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on stromal cells of Human stomach 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.

  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/5000 dilution

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    There was an additional band detected in the heart lysate.

  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution

    Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : Raji (Human Burkitt's lymphoma 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?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution

    Lane 1 : Mouse brain lysate at 10 µg
    Lane 2 : Mouse heart lysate at 10 µg
    Lane 3 : Mouse spleen lysate at 10 µg
    Lane 4 : Mouse kidney lysate at 10 µg
    Lane 5 : Rat brain lysate at 10 µg
    Lane 6 : Rat heart lysate at 10 µg
    Lane 7 : Rat kidney lysate at 10 µg
    Lane 8 : Rat spleen lysate at 10 µg
    Lane 9 : C6 (Rat glial tumor cell line) whole cell lysate at 20 µg
    Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
    Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    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 1,2 and 3: 15 seconds; Lane 4: 3 minutes; Lane 5,6,7 and 8: 15 seconds; Lane 9 and 10: 5 seconds; Lane 11: 3 minutes.

    There was an additional band detected in the heart lysate.

  • Immunoprecipitation - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Immunoprecipitation - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

    Annexin-6/ANXA6 was immunoprecipitated from 1mg of Human fetal liver whole cell lysate with ab201023 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab201023 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: ab201023 IP in Human fetal liver whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control - (ab172730) instead of ab201023 in Human fetal liver whole cell lysate.

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

    Exposure time: 1 second.

  • Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
    Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (0)

Publishing research using ab201023? Please let us know so that we can cite the reference in this datasheet.

ab201023 has not yet been referenced specifically in any publications.

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