Anti-Annexin-2/ANXA2 antibody [RM1229]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (human prostate carcinoma epithelial cell Left) / MCF7 (human breast adenocarcinoma epithelial cell. Right) cells labelling Annexin-2/ANXA2 with ab321903 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : LNCaP

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunohistochemical analysis of paraffin-embedded Human cervical squamous carcinoma tissue labeling Annexin-2/ANXA2 with ab321903 at 1/2000 (0.252 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cervical squamous carcinoma. The section was incubated with ab321903 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Annexin-2/ANXA2 with ab321903 at 1/2000 (0.252 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney. The section was incubated with ab321903 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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunofluorescent analysis of 100% methanol-fixed 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Annexin-2/ANXA2 with ab321903 at 1/1000 (0.504 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in MCF7 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : LNCaP.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Annexin-2/ANXA2 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab321903 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab321903 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab321903 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab321903 in MCF7 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/30 dilution

All lanes:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 10s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunofluorescent analysis of 100% methanol-fixed 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Annexin-2/ANXA2 with ab321903 at 1/1000 (0.504 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in NIH/3T3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : A20.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Annexin-2/ANXA2 with ab321903 at 1/2000 (0.252 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney. The section was incubated with ab321903 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Annexin-2/ANXA2 with ab321903 at 1/2000 (0.252 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat kidney. The section was incubated with ab321903 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)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A20 (mouse reticulum sarcoma B lymphocyte Left) / NIH/3T3 (mouse embryonic fibroblast Right) cells labelling Annexin-2/ANXA2 with ab321903 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : A20

• IP

Supplier Data

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Annexin-2/ANXA2 was immunoprecipitated from 0.35 mg PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate with ab321903 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab321903 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 2 : ab321903 IP in PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab321903 in PC-12 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/30 dilution

All lanes:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• IP

Supplier Data

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

Annexin-2/ANXA2 was immunoprecipitated from 0.35 mg 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate with ab321903 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab321903 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate
Lane 2 : ab321903 IP in 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab321903 in 4T1 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/30 dilution

All lanes:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

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

Exposure time : Lane 1 : 81 seconds Lane 2 : 37 seconds Lane 3-4 : 15 seconds

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Human spleen 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: 36 kDa

false

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

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

Low expression : LNCaP, Jurkat, 293T.

The molecular weight observed is consistent with what has been described in the literature (PMID : 16373665 and PMID : 22359590).

Anti-Annexin-2/ANXA2 antibody (ab321903, Left) staining at 1/1000 dilution showed similar expression profile/ molecular weight with Anti-Annexin-2/ANXA2 antibody (ab41803, Right) at 1/1000 dilution.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 8:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 9:

4T1 (mouse mammary gland carcinoma epithelial cell) 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: 36 kDa,124 kDa,33 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

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

Exposure time : Lane 1 : 180 seconds Lane 2-4 : 37 seconds

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen 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: 36 kDa

false

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

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

Exposure time : Lane 1 : 180 seconds Lane 2-4 : 37 seconds

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Lane 4:

Rat spleen 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

false

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (AB321903)

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

Low expression : A20.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-4 : 15 seconds Lane 5 : 180 seconds

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody [RM1229] (ab321903) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

A20 (mouse reticulum sarcoma B lymphocyte) 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

false