Recombinant
RabMAb

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

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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

    Suitable for: Flow Cyt, IP, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Mouse Annexin-2/ANXA2 aa 1-300. The exact sequence is proprietary.
    Database link: P07356

  • 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

Applications

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.

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.

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.
  • Information by UniProt

  • Database links

    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.

References

ab189473 has not yet been referenced specifically in any publications.

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

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