Anti-Annexin V/ANXA5 antibody [EPR28047-61] KO Tested (ab313581)

Knockout Tested Rabbit Recombinant Monoclonal Annexin V/ANXA5 antibody. Suitable for WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Recombinant fragment - Human samples.

Be the first to review this product! Submit a review

Images

Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (AB313581), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	Flow Cyt (Intra)	IHC-P	IP	ICC/IF	IHC-Fr
Human	Tested	Tested	Tested	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Tested	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Recombinant fragment - Human	Tested	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Recombinant fragment - Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Recombinant fragment - Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Recombinant fragment - Human, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Recombinant fragment - Human, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Recombinant fragment - Human, Rat	Dilution info -	Notes -

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

See full target information Anxa5

Function

This protein is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade.

Alternative names

Anx5, Anxa5, Annexin A5, Anchorin CII, Annexin V, Annexin-5, Calphobindin I, Endonexin II, Lipocortin V, Placental anticoagulant protein 4, Placental anticoagulant protein I, Thromboplastin inhibitor, Vascular anticoagulant-alpha, CPB-I, PP4, PAP-I, VAC-alpha

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal Annexin V/ANXA5 antibody. Suitable for WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Recombinant fragment - Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR28047-61
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Annexin V also known as ANXA5 is a protein with a mass of approximately 35-36 kDa. This protein belongs to the annexin family which includes proteins capable of binding to phospholipids in a calcium-dependent manner. Annexin V is mostly expressed in placental tissue though it can be found in various other tissues like the vascular endothelium. Researchers often utilize annexin V for staining dead cells or identifying apoptosis using assays such as annexin V FITC or 7-AAD staining. It forms part of the annexin V binding buffer which enhances its binding affinity.

Biological function summary

Annexin V has an important role in cell membrane dynamics and signaling. The protein does not function as part of a larger complex but its ability to bind negatively charged phospholipids like phosphatidylserine makes it important in apoptosis. During apoptosis phosphatidylserine translocates to the outer leaflet of the plasma membrane where annexin V marks these apoptotic cells. This property enables annexin V-based assays to detect early stages of apoptosis.

Pathways

Annexin V involvement is seen in pathways associated with blood coagulation and apoptosis. In the apoptosis pathway annexin V interacts with apoptotic markers to identify dying cells. It relates to proteins such as caspases which are key executors of apoptosis. In coagulation annexin V has been shown to interact with prothrombinase complex and can affect calcium ion concentration thereby influencing blood clotting processes.

Associated diseases and disorders

Annexin V plays a role in antiphospholipid syndrome and cardiovascular diseases. Annexin V has been researched for its connection to antiphospholipid antibodies where it affects normal annexin V function impacting clot formation. In cardiovascular diseases annexin V can interact with proteins such as thrombomodulin influencing thrombotic and atherosclerotic pathways. Understanding these interactions helps researchers develop potential therapeutic approaches for related conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Annexin V/ANXA5 Western blot staining using rabbit Anti-Annexin V/ANXA5 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Performed under reducing conditions.
In Western blot, ab313581 was shown to bind specifically to ANXA5. A band was observed at 35 kDa in wild-type HAP1 cell lysates whereas loss of signal was observed in the ANXA5 knockout cell line.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581) at 1/1000 dilution
Lane 1: Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg
Lane 2: ANXA5 knockout HAP1 whole cell lysate at 20 µg
Lane 3: Human liver tissue lysate at 20 µg
Lane 4: Human testis tissue lysate at 20 µg
Lane 5: Mouse liver tissue lysate at 20 µg
Lane 6: Mouse testis tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 35 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded (A) Liver tissue from wild-type C57BL/6J mice and (B) Liver tissue from ANXA5 knockout mice tissue labelling ANXA5 with ab313581 at 1/10000 (0.049 μg/ml) followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) liver tissue from wild-type C57BL/6J mice and no staining on (B) liver tissue from ANXA5 knockout mice. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Annexin A5-KO homozygous mice (Strain ID: T028565).
Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM/TBST.

The identity of the bands above 50 kDa is unknown.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a loading control.
All lanes: Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581) at 1/1000 dilution
Lane 1: Wild-type mouse liver tissue lysate at 20 µg
Lane 2: Anxa5 knockout mouse liver tissue lysate at 20 µg
Lane 3: Wild-type mouse lung tissue lysate at 20 µg
Lane 4: Anxa5 knockout mouse lung tissue lysate at 20 µg
Lane 5: Wild-type mouse heart tissue lysate at 20 µg
Lane 6: Anxa5 knockout mouse heart tissue lysate at 20 µg
Lane 7: Wild-type mouse kidney tissue lysate at 20 µg
Lane 8: Anxa5 knockout mouse kidney tissue lysate at 20 µg
Lane 9: Wild-type mouse colon tissue lysate at 20 µg
Lane 10: Anxa5 knockout mouse colon tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 26 kDa
Exposure time: 35s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded (A) Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet and (B) ANXA5 knockout HAP1 cell pellet tissue labelling ANXA5 with ab313581 at 1/10000 (0.049 μg/ml) followed by a ready to use secondary antibody LeicaDS9800 (Bond^™ Polymer Refine Detection). Positive staining on (A) Wild-type HAP1 cell pellet and negative staining on (B) ANXA5 knockout HAP1 cell pellet. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.
Exposure time: 26 seconds
All lanes: Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581) at 1/1000 dilution
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 35 kDa
Exposure time: 26s
Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody does not cross-react with human ANXA4.
In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution
Exposure time: 180 seconds
All lanes: Western blot - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581) at 1/1000 dilution
Lane 1: His-tagged human ANXA4 recombinant protein at 10 ng
Lane 2: His-tagged human ANXA5 recombinant protein at 10 ng
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 35 kDa
Exposure time: 180s
Flow Cytometry (Intracellular) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Annexin V/ANXA5 with ab313581 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized ANXA5 KO HAP1 (human ANXA5 knockout chronic myelogenous leukemia near-haploid cell, Left) / parental HAP1 (Right) cells labelling Annexin V/ANXA5 with ab313581 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Annexin V/ANXA5 with ab313581 at 1/10000 (0.049 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Annexin V/ANXA5 with ab313581 at 1/10000 (0.049 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse liver. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Annexin V/ANXA5 with ab313581 at 1/10000 (0.049 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded Human prostatic hype tissue labeling Annexin V/ANXA5 with ab313581 at 1/10000 (0.049 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human prostatic hyperplasia. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR28047-61] (ab313581)
Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Annexin V/ANXA5 with ab313581 at 1/10000 (0.049 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human testis. The section was incubated with ab313581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins