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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunohistochemistry [1]
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- Product number
- PA5-19711 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Annexin A2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Heat mediated antigen retrieval recommended prior to tissue staining.
- Concentration
- 0.7 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HeLa Whole Cell Lysate using Product # PA5-19711, Annexin II primary antibody at a dilution of 1 µg/mL (lane 1). Staining of HEK293 Whole Cell Lysate at a dilution of 1 µg/mL (lane 2). Blot treated with a secondary HRP-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:3000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Annexin A2 was achieved by transfecting HeLa with Annexin A2 specific siRNAs (Silencer® select Product # S1384, S1385). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Annexin A2 untransfected cells (lane 1), non-targeting scrambled siRNA transfected cells (lane 2) and knockdown cells (lane 3). The blot was probed with Annexin A2 Polyclonal Antibody (Product # PA5-19711, 1:7250 dilution ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Annexin A2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Annexin A2 Polyclonal Antibody(Product # PA5-19711) and a 40kDa band corresponding to Annexin A2 was observed across cell lines and tissues tested. Whole cell extracts (40 µg lysate) of A549 (Lane 1), HCT 116 (Lane 2), HEK-293 (Lane 3), HeLa (Lane 4), Mouse Kidney (Lane 5), Rat Kidney (Lane 6), Mouse Testis (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical (formalin-fixed, paraffin-embedded) staining of Human Breast Carcinoma tissue using Product # PA5-19711, anti-Annexin II antibody. Primary antibody was used at a concentration of 1 µg/mL and exposed for 15 mins at room temp. The sample was pretreated using heat mediated antigen retrieval with Sodium Citrate Buffer (pH6/20mins). The detection method was a HRP conjugated polymer, DAB chromogen and the sample was counterstained with haematoxylin and mounted with DPX.
