Recombinant Anti-Annexin-2/ANXA2 antibody [EPR18157-104] (ab189473)

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 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

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).

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.

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.

Intracellular 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 Fluorr^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

 

 

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.