Name: Anti-Annexin A1/ANXA1 antibody [EPR19342]
Brand: RabMAb
SKU: ab214486

Product name

Anti-Annexin A1/ANXA1 antibody [EPR19342]
See all Annexin A1/ANXA1 primary antibodies
Description

Rabbit monoclonal [EPR19342] to Annexin A1/ANXA1
Host species

Rabbit
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/ANXA1 aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P04083

Run BLAST with Run BLAST with
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.

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

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19342
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Annexin A1/ANXA1 protein (ab86446)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

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.

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.
Target information above from: UniProt accession P04083 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 301 Human
- Entrez Gene: 16952 Mouse
- Entrez Gene: 25380 Rat
- Omim: 151690 Human
- SwissProt: P04083 Human
- SwissProt: P10107 Mouse
- SwissProt: P07150 Rat
- Unigene: 494173 Human
- Unigene: 248360 Mouse
- Unigene: 1792 Rat
see all
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

Western blot - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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 Cytometry - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunoprecipitation - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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 Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 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.
Western blot - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

All lanes : Anti-Annexin A1/ANXA1 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
All lanes : 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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunocytochemistry/ Immunofluorescence - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Western blot - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

Lanes 1-2 : Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486) at 1/5000 dilution
Lanes 3-7 : Anti-Annexin A1/ANXA1 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
All lanes : 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Western blot - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

Lanes 1-2 : Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486) at 1/5000 dilution
Lane 3 : Anti-Annexin A1/ANXA1 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
All lanes : 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

This product has been referenced in:

Feng J et al. Silencing of Annexin A1 suppressed the apoptosis and inflammatory response of preeclampsia rat trophoblasts. Int J Mol Med 42:3125-3134 (2018). Read more (PubMed: 30272262) »
Su D et al. PTIP chromatin regulator controls development and activation of B cell subsets to license humoral immunity in mice. Proc Natl Acad Sci U S A 114:E9328-E9337 (2017). Flow Cyt ; Mouse . Read more (PubMed: 29078319) »

See all 2 Publications for this product

Publishing research using ab214486? Please let us know so that we can cite the reference in this datasheet.

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Recombinant Anti-Annexin A1/ANXA1 antibody [EPR19342] (ab214486)

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Annexin A1 antibody [EPR19342]