PA5-37474

antibody from Invitrogen Antibodies

Targeting: ANXA2 ANX2, ANX2L4, CAL1H, LIP2, LPC2D

Antibody data

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Validation data

Product number: PA5-37474 - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Annexin A2 (Ser26) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: A suggested positive control for Western blot is HepG2 or JK cells. The homology between synthetic peptide sequence and Mouse/Rat is 100%.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: -20°C

ANXA2 protein structure - PA5-37474 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 2 FIGURE PKC phosphorylates ANXA2 on Ser25 side chain. (a) Purified recombinant GST, GST-ANXA2, and GST-TIMAP proteins were incubated with or without active PKCalpha for 30 min. Amount of loaded proteins were verified by SDS-PAGE followed by Coomassie staining. Phosphorylation of proteins was analyzed by western blot using anti-phosphoSer PKC substrate and phospho-Ser25 annexin A2-specific antibodies. (b) Pull down of endothelial cell lysate was made using GST, GST-tagged TIMAP wild type, S331D or S331D recombinant full-length TIMAP proteins as baits. Samples were tested for ANXA2 in western blot experiment. (c) Total cell lysates of control, nonsilencing RNA and PKC-specific siRNA transfected HPAEC treated with PMA (1 muM for 30 min) or PMA (1 muM for 30 min) after a pretreatment with Go6983 (1 muM for 30 min) were analyzed by western blot using PKCalpha, actin, ANXA2, and phospho-Ser25 ANXA2-specific antibodies. (d) Western blot analysis of control, PMA (1 muM, 30 min), PMA (1 muM, 30 min) with Go6983 pretreatment (1 muM, 30 min), okadaic acid (5 nM, 30 min), tautomycetin (1 muM, 30 min), nonsiRNA, or TIMAP-specific siRNA-treated BPAEC cells are shown. Phospho-Ser25 ANXA2 level was normalized to ANXA2 protein level. Densitometric analysis of western blots are shown as mean +- SD ( n = 3). Statistical analysis was done with ANOVA and marked by asterisks; *** p < .001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Phosphorylation of AnxA2 in breast cancer cells. ( A ) The expression of pAnxA2-S25, pAnxA2-Y23, and AnxA2 were analyzed in non-tumorigenic mammary epithelial cells and TNBC cells by immunoblot analysis. The membrane was stripped and reprobed with anti-beta-actin antibody for loading control. ( B ) Bar chart showing the densitometric analysis of pAnxA2-S25, pAnxA2-Y23 and AnxA2 bands of the immunoblot of panel A. Intensity of each bands were normalized by loading control. Each bar represents the mean +- SE of three independent experiments. ( C ) The HCC1395 cells were transfected with a plasmid vector expressing GFP alone, AnxA2-WT-GFP, and AnxA2-Y23F-GFP. The expression of pAnxA2-Y23, AnxA2 and GFP were analyzed by immunoblot analysis. The membrane was stripped and reprobed with anti-beta-actin antibody for loading control. ( D ) The HCC1395 transfected cells (6 x 10 5 ) were plated in 100mm Petri dish for overnight and then switched to serum free medium. After 24 h, the AnxA2-GFP complex was immunoprecipitated from the medium using anti-GFP antibody (Catalog no. DSHB-GFP-12A6; DSHB) and expression of AnxA2 secretion was analyzed by anti-AnxA2 antibody using immunoblot analysis. Uncropped Western Blots of ( A , C , D ) are available in Figure S3 .