Human/Mouse/Rat Annexin A2 Antibody

Detection of Human/Mouse/Rat Annexin A2 by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat Annexin A2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3928) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Annexin A2 at approximately 39 kDa (as indicated). This experiment was conducted using Immunoblot Buffer Group 2.

Detection of Human and Mouse Annexin A2 by Simple Western^TM. Simple Western shows lysates of Exosome Standards (HT‑29) (NBP3-11685) and COLO 205 human colorectal adenocarcinoma cell line, loaded at 0.5 mg/ml. A specific band was detected for Annexin A2 at approximately 44 kDa (as indicated) using 100 µg/mL of Goat Anti-Human/Mouse/Rat Annexin A2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3928). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Annexin A2 in Human Liver. Annexin A2 was detected in immersion fixed paraffin-embedded sections of normal human liver using Goat Anti-Human/Mouse/Rat Annexin A2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3928) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membranes of hepatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot Shows Human Annexin A2 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Annexin A2 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat Annexin A2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3928) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Annexin A2 at approximately 37 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse Annexin A2 by Western Blot MIPs induce Src-dependent tyrosine phosphorylation and translocation of A2 to the cell surface. A ARPE-19 cells were incubated for 18 h with basic medium (Basic), activated THP-1 cell conditioned medium (CM), or THP-1 CM with RPE CM with or without addition of anti-human MIP-1 alpha / beta or non-immune IgG. Cell surface A2 (sA2) was labeled using membrane impermeable biotinylation and captured with NeutrAvidin agarose beads. Labeled cell surface proteins were resolved by SDS-PAGE and cell surface (sA2) and total cell lysate A2 (tA2) probed by immunoblot with mouse (BD #610069) or rabbit (Cell Signaling #8235) anti-A2 IgG. b Pixel density of immunoblotted bands from 3-5 separate experiments were quantified and normalized to the unstimulated sample (Basic). c ARPE-19 cells were incubated with basic media with or without recombinant human MIP-1 alpha / beta (MIPs, 200 ng/ml each), and with or without the Src kinase inhibitor PP2 or its control PP3 (10 µM, 3 h). Cell lysates were probed for pY23-A2, tA2, pY416 activated Src, total Src (tSrc), and GAPDH, as a loading control. d Pixel density of immunoblotted bands from 3-6 separate experiments were quantified and normalized to the unstimulated sample (Basic). e ARPE-19 cells were incubated with or without recombinant human MIP-1 alpha / beta (MIPs, 200 ng/ml each) in the presence or absence of PP2 or PP3, as in (c). f Pixel density of immunoblotted bands from 3–7 separate experiments were quantified and normalized to the unstimulated sample (Basic). Shown for (b, d, f) are mean values ± SE, with p values determined by unpaired one-way ANOVA and the post hoc Tukey test. Source data for (b, d, and f) are provided as a Source File. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39384746), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Annexin A2 by Flow Cytometry Effect of anti-annexin A2 IgG pretreatment on dispase-induced PVR.a, b Wild type C57Bl/6 mice were pretreated with an anterior chamber injection of anti-A2 IgG (1A7 or 2E6), control IgG (1D4), or PBS, two days prior to intravitreal dispase injection. At 2 (a) and 4 (b) weeks, the eyes were harvested, fixed, embedded, sectioned, and stained with H and E. Scale bars for (a and b) are 200 um. PVR histology (c) and RPE migration (d) scores were recorded by trained, masked observers. Shown for (c and d) are mean ± SE, n = 7 animals/group. p values were determined by unpaired one-way ANOVA and the post hoc Tukey test. Source data for (c and d), are provided as a Source File. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39384746), licensed under a CC-BY license. Not internally tested by R&D Systems.