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Signal Transduction Cytoskeleton / ECM Cell Adhesion Annexins
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RecombinantRabMAb

Recombinant Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

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  • Certificate of Compliance
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Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Flow Cytometry - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
  • Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18157-104] to Annexin-2/ANXA2
  • Suitable for: Flow Cyt, IP, WB, IHC-P
  • Reacts with: Mouse, Rat, Human

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Extraction Buffer 5X PTR (ab193970)

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Overview

  • Product name

    Anti-Annexin-2/ANXA2 antibody [EPR18157-104]
    See all Annexin-2/ANXA2 primary antibodies
  • Description

    Rabbit monoclonal [EPR18157-104] to Annexin-2/ANXA2
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Mouse
    IP
    Mouse
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

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

  • Positive control

    • WB: Human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; HeLa, K562, C6 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Rat spleen lysate. IHC-P: Mouse liver tissue. Flow Cyt: HeLa cells. IP: NIH/3T3 whole cell lysate.
  • General notes

     This product was previously labelled as Annexin A2

     

    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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18157-104
  • 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-2/ANXA2 antibody [EPR18157-104] - BSA and Azide free (ab227486)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Annexin-2/ANXA2 protein (ab93005)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab189473 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Mouse
IP
Mouse
WB
Mouse
Rat
Human
Application Abreviews Notes
Flow Cyt
1/500.
IP
1/30.
WB
1/5000. Detects a band of approximately 36, 34 kDa (predicted molecular weight: 38 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is not recommended in human or rat due to nonspecific or negative staining.

Notes
Flow Cyt
1/500.
IP
1/30.
WB
1/5000. Detects a band of approximately 36, 34 kDa (predicted molecular weight: 38 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is not recommended in human or rat due to nonspecific or negative staining.

Target

  • Function

    Calcium-regulated membrane-binding protein whose affinity for calcium is greatly enhanced by anionic phospholipids. It binds two calcium ions with high affinity. May be involved in heat-stress response.
  • Sequence similarities

    Belongs to the annexin family.
    Contains 4 annexin repeats.
  • Domain

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

    Phosphorylation of Tyr-24 enhances heat stress-induced translocation to the cell surface.
    ISGylated.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix > basement membrane. Melanosome. In the lamina beneath the plasma membrane. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Translocated from the cytoplasm to the cell surface through a Golgi-independent mechanism.
  • Target information above from: UniProt accession P07355 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 302 Human
    • Entrez Gene: 12306 Mouse
    • Entrez Gene: 56611 Rat
    • Omim: 151740 Human
    • SwissProt: P07355 Human
    • SwissProt: P07356 Mouse
    • SwissProt: Q07936 Rat
    • Unigene: 511605 Human
    • Unigene: 238343 Mouse
    • Unigene: 90546 Rat
    see all
  • Alternative names

    • Annexin A2 antibody
    • Annexin II antibody
    • Annexin II, heavy chain antibody
    • Annexin-2 antibody
    • ANX 2 antibody
    • ANX2 antibody
    • ANX2L4 antibody
    • ANXA2 antibody
    • ANXA2_HUMAN antibody
    • arylsulfatase B antibody
    • CAL1H antibody
    • Calpactin I heavy chain antibody
    • calpactin I heavy polypeptide (p36) antibody
    • Calpactin I heavy polypeptide antibody
    • Calpactin-1 heavy chain antibody
    • chromobindin 8 antibody
    • Chromobindin-8 antibody
    • Epididymis secretory protein Li 270 antibody
    • HEL S 270 antibody
    • LIP2 antibody
    • Lipocortin II antibody
    • LPC2 antibody
    • LPC2D antibody
    • p36 antibody
    • P36 protein antibody
    • PAP-IV antibody
    • Placental anticoagulant protein IV antibody
    • Protein I antibody
    see all

Images

  • Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    All lanes : Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473) at 1/5000 dilution

    Lane 1 : Human fetal brain 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 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 36 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    All lanes : Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473) at 1/5000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 3 : C6 (rat glial tumor cell line) whole cell lysate
    Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 34,36 kDa why is the actual band size different from the predicted?



    Exposure times: Lane 1: 3 seconds; Lane 2,4: 1 second; Lane 3: 30 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 16373665).

  • Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Western blot - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    All lanes : Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 36 kDa why is the actual band size different from the predicted?



    Exposure times: Lane 1: 2 seconds; Lanes 2-4: 1 second; Lane 5: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Annexin-2/ANXA2 with ab189473 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the sinusoid of mouse liver (PMID: 23188673). 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.

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

  • Flow Cytometry - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Flow Cytometry - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Annexin-2/ANXA2 with ab189473 at 1/500 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.

  • Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

    Annexin-2/ANXA2 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab189473 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab189473 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 µg (Input). 

    Lane 2: ab189473 IP in NIH/3T3 whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189473 in NIH/3T3 whole cell lysate.

    Exposure time: 1 second.

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

  • Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)
    Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

    • Datasheet

    Certificate of Compliance

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

  • References (0)

    Publishing research using ab189473? Please let us know so that we can cite the reference in this datasheet.

    ab189473 has not yet been referenced specifically in any publications.

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