Annexin V Antibody (38701) [Unconjugated]

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Annexin V Monoclonal Antibody (Catalog # MAB3991) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Annexin V at approximately 32 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Annexin V was detected in immersion fixed paraffin-embedded sections of human placenta using Mouse Anti-Human Annexin V Monoclonal Antibody (Catalog # MAB3991) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to syncytiotrophoblasts. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of human lung tissue, loaded at 0.2 mg/mL. A specific band was detected for Annexin V at approximately 39 kDa (as indicated) using 2.5 µg/mL of Mouse Anti-Human Annexin V Monoclonal Antibody (Catalog # MAB3991). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Simple Western lane view shows lysates of A549 human lung carcinoma cell line exosomes, HT‑29 human colon adenocarcinoma cell line exosomes, and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Annexin V at approximately 38 kDa (as indicated) using 50 µg/mL of Mouse Anti-Human Annexin V Monoclonal Antibody (Catalog # MAB3991) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Annexin V knockout HEK293T cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Annexin V Monoclonal Antibody (Catalog # MAB3991) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Annexin V at approximately 32 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Extracellular vesicle secretion induced by Sorafenib and their miRNA content. (A) Particle size and concentration analysis of Large, Small, and Very Small EVs obtained at 6 and 24 h. (B) Expression of EV markers and cellular contaminants in Large, Small, and Very Small EVs and cell lysates obtained at 24 h after Sorafenib treatment. Total lane protein content was used as the loading control. (C) Representative images of cryo-EM of Small and Very Small EVs obtained at 6 and 24 h from the control and Sorafenib-treated cells. (D) Assessment of EV size (nm) in the cryo-EM images. (E) Number of EVs quantified in the cryo-EM images. (F) miRNA expression in the three fractions of EVs at 6 h. (G) miRNA expression in the three fractions of EVs at 24 h. Fold-change values were calculated between Sorafenib and the control treated samples. Results are expressed as the mean ± SEM of six independent experiments. Ns, non-significant; * p ≤0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001 between the miRNA expression in the control and Sorafenib derived EVs. Multiple comparison test statistics are expressed with lower case a–i. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36078082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Extracellular vesicle secretion induced by Sorafenib and their miRNA content. (A) Particle size and concentration analysis of Large, Small, and Very Small EVs obtained at 6 and 24 h. (B) Expression of EV markers and cellular contaminants in Large, Small, and Very Small EVs and cell lysates obtained at 24 h after Sorafenib treatment. Total lane protein content was used as the loading control. (C) Representative images of cryo-EM of Small and Very Small EVs obtained at 6 and 24 h from the control and Sorafenib-treated cells. (D) Assessment of EV size (nm) in the cryo-EM images. (E) Number of EVs quantified in the cryo-EM images. (F) miRNA expression in the three fractions of EVs at 6 h. (G) miRNA expression in the three fractions of EVs at 24 h. Fold-change values were calculated between Sorafenib and the control treated samples. Results are expressed as the mean ± SEM of six independent experiments. Ns, non-significant; * p ≤0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001 between the miRNA expression in the control and Sorafenib derived EVs. Multiple comparison test statistics are expressed with lower case a–i. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36078082), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, KO
• Clone: 38701
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Annexin V Antibody (38701) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Annexin V
  Met1-Asp320
  Accession # P08758
• Specificity: Detects human Annexin V in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-25 ug/mL
  • Knockout Validated
  • Simple Western 2.5-50 ug/mL
  • Western Blot 0.25 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Annexin V Antibody (38701) [Unconjugated]

• Anchorin CII
• annexin A5
• Annexin V
• annexin-5
• ANX5
• ANXA5
• ANXV
• Calphobindin I
• Endonexin II
• ENX2CBP-I
• Lipocortin V
• PAP-I
• Placental anticoagulant protein 4
• Placental anticoagulant protein I
• PP4
• Thromboplastin inhibitor
• VAC-alpha
• Vascular anticoagulant-alpha

Background

Annexin V is a member of a calcium and phospholipid binding family of proteins with vascular anticoagulant activity. It is also known as PP4, PAP I, CPB-I, CaBP33, VACa, anchorin CII, lipocortin-V, endonexin II, and thromboplastin inhibitor.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Annexin V Antibody (MAB3991)(1)

Publication using MAB3991

• Mukhopadhyay, U;Levantovsky, S;Carusone, TM;Gharbi, S;Stein, F;Behrends, C;Bhogaraju, S; A ubiquitin-specific, proximity-based labeling approach for the identification of ubiquitin ligase substrates Science advances 2024-08-09 [PMID: 39121224] (Western Blot, Human)

Bioinformatics

• Uniprot:
  • P08758()