Recombinant 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
Overview
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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 HumanIHC-P MouseIP MouseWB MouseRatHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18157-104 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
|
IHC-P |
Mouse
|
IP |
Mouse
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WB |
Mouse
Rat
Human
|
Application | Abreviews | Notes |
---|---|---|
Flow Cyt |
1/500.
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|
IP |
1/30.
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|
WB |
1/5000. Detects a band of approximately 36, 34 kDa (predicted molecular weight: 38 kDa).
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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 |
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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
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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
modificationsPhosphorylation 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
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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
see all -
Alternative names
- Annexin A2 antibody
- Annexin II antibody
- Annexin II, heavy chain antibody
see all
Images
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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).
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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.
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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.
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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.
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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.
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.
Datasheets and documents
Certificate of Compliance
References (0)
ab189473 has not yet been referenced specifically in any publications.