Anti-Annexin V/ANXA5 antibody [EPR28047-61]

• IHC-P

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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

• IHC-P

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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.

• IHC-P

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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

• IHC-P

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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

• Flow Cyt (Intra)

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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 (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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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

• IHC-P

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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).

• IHC-P

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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

• Flow Cyt (Intra)

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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 (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

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

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] (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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa

false

Exposure time: 180s

• WB

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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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa

false

Exposure time: 26s

• WB

Supplier Data

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. 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa

false

Exposure time: 35s

• WB

Supplier Data

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 (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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa

false

Exposure time: 180s