Validated using a knockout cell line
Recombinant
RabMAb

Anti-Annexin A1 antibody [EPR19342] (ab214486)

Overview

  • Product name
    Anti-Annexin A1 antibody [EPR19342]
    See all Annexin A1 primary antibodies
  • Description
    Rabbit monoclonal [EPR19342] to Annexin A1
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. within Human Annexin A1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P04083

  • Positive control
    • WB: K562, BxPC-3, C2C12, C6, PC-12 and NIH/3T3 whole cell lysates; human fetal brain, fetal kidney and placenta lysates; mouse and rat kidney and spleen lysates. IHC-P: Human tonsil, breast, endometrial cancer and bladder cancer tissues; rat lung tissue; mouse spleen tissue. ICC/IF: BxPC-3 and NIH/3T3 cells. Flow Cyt: NIH/3T3 cells. IP: K562 whole cell lysate.
  • General notes

    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 ab214486 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 1/2000. Detects a band of approximately 37, 33 kDa (predicted molecular weight: 38 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/600.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30.

Target

  • Function
    Calcium/phospholipid-binding protein which promotes membrane fusion and is involved in exocytosis. This protein regulates phospholipase A2 activity. It seems to bind from two to four calcium ions with high affinity.
  • 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
    Phosphorylated by protein kinase C, epidermal growth factor receptor/kinase and TRPM7. Phosphorylation results in loss of the inhibitory activity.
  • Cellular localization
    Nucleus. Cytoplasm. Cell projection > cilium. Basolateral cell membrane. Found in the cilium, nucleus and basolateral cell membrane of ciliated cells in the tracheal endothelium (By similarity). Found in the cytoplasm of type II pneumocytes and alveolar macrophages.
  • Information by UniProt
  • Database links
  • Alternative names
    • Annexin 1 antibody
    • Annexin A1 antibody
    • Annexin I (lipocortin I) antibody
    • Annexin I antibody
    • Annexin-1 antibody
    • AnnexinA1 antibody
    • AnnexinI antibody
    • ANX 1 antibody
    • ANX A1 antibody
    • ANX1 antibody
    • ANXA 1 antibody
    • ANXA1 antibody
    • ANXA1 protein antibody
    • ANXA1_HUMAN antibody
    • Calpactin 2 antibody
    • Calpactin II antibody
    • Calpactin-2 antibody
    • CalpactinII antibody
    • Chromobindin 9 antibody
    • Chromobindin-9 antibody
    • Chromobindin9 antibody
    • HGNC:533 antibody
    • Lipocortin 1 antibody
    • Lipocortin I antibody
    • Lipocortin1 antibody
    • LipocortinI antibody
    • LPC 1 antibody
    • LPC1 antibody
    • p35 antibody
    • Phospholipase A2 inhibitory protein antibody
    see all

Images



  • Predicted band size : 38 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Annexin A1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab214486 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab214486 was shown to specifically react with Annexin A1 when Annexin A1 knockout samples were used. Wild-type and Annexin A1 knockout samples were subjected to SDS-PAGE. Ab214486 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

    Nuclear and cytoplasmic staining on human tonsil tissue is observed [PMID:9720986].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized BxPC-3 (Human pancreas adenocarcinoma cell line) cells labeling Annexin A1 with ab214486 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing membrane and weak cytoplasmic staining on BxPC-3 cell line.

    The nuclear counterstain is DAPI (blue).

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

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Annexin A1 with ab214486 at 1/600 dilution (red) compared with 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 A1 was immunoprecipitated from 0.35 mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab214486 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab214486 at 1/1000 dilution.

    VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: K562 whole cell lysate 10µg (Input).

    Lane 2: ab214486 IP in K562 whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214486 in K562 whole cell lysate.

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

    Exposure time: 1 second.

  • All lanes : Anti-Annexin A1 antibody [EPR19342] (ab214486) at 1/10000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 2 : BxPC-3 (Human pancreas adenocarcinoma cell line) whole cell lysate
    Lane 3 : C2C12 (Mouse myoblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size : 38 kDa
    Observed band size : 33,37 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 second

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The two band 37kDa is the full length band and the 33kDa supposed to be the cleavage form, this expression pattern observed is consistent with what has been described in the literature (PMID: 25510623 and 20679535).

  • Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic staining on human breast tissue is observed [PMID:16949910].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Annexin A1 with ab214486 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing membrane and cytoplasmic staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

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

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Lanes 1 - 2 : Anti-Annexin A1 antibody [EPR19342] (ab214486) at 1/5000 dilution
    Lanes 3 - 7 : Anti-Annexin A1 antibody [EPR19342] (ab214486) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 4 : Mouse kidney lysate
    Lane 5 : Mouse spleen lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size : 38 kDa
    Observed band size : 33,37 kDa (why is the actual band size different from the predicted?)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2/6/7: 1 seconds; Lane 3: 15 seconds; Lane 4/5: 30 seconds.

  • Immunohistochemical analysis of paraffin-embedded human endometrial cancer tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic and weak membrane staining on human endometrial cancer tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Lanes 1 - 2 : Anti-Annexin A1 antibody [EPR19342] (ab214486) at 1/5000 dilution
    Lane 3 : Anti-Annexin A1 antibody [EPR19342] (ab214486) at 1/20000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human placenta lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size : 38 kDa
    Observed band size : 33,37 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: 15 seconds; Lane 3: 1 second.

  • Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear, cytoplasmic and membrane staining on human bladder cancer tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic staining on rat lung tissue is observed [PMID:15133855] [PMID:9720986].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Annexin A1 with ab214486 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic staining on mouse spleen tissue is observed [PMID:9720986].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

References

ab214486 has not yet been referenced specifically in any publications.

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