Anti-Annexin-2/ANXA2 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Annexin-2/ANXA2 antibody (AB41803)

ab41803 staining Annexin-2 in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab41803 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Annexin-2/ANXA2 antibody (AB41803)

ICC/IF image of ab41803 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab41803, 1μg/ml) overnight at +4°C. The secondary antibody (green) was DyLight^® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-2/ANXA2 antibody (AB41803)

IHC image of Annexin-2/ANXA2 staining in FFPE human breast carcinoma FFPE section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab41803, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• WB

Supplier Data

Western blot - Anti-Annexin-2/ANXA2 antibody (AB41803)

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] (<a href='/en-us/products/primary-antibodies/annexin-2-anxa2-antibody-rm1229-ab321903'>ab321903</a>) 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

Unknown

Western blot - Anti-Annexin-2/ANXA2 antibody (AB41803)

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody (ab41803) at 1 µg/mL

All lanes:

Western blot - Recombinant Human Annexin-2/ANXA2 protein (6x His tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-annexin-2-anxa2-protein-ab93005'>ab93005</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 38 kDa

true

Exposure time: 20s

• WB

Project

Western blot - Anti-Annexin-2/ANXA2 antibody (AB41803)

All lanes:

Western blot - Anti-Annexin-2/ANXA2 antibody (ab41803) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 38 kDa

Observed band size: 38 kDa

false