Apolipoprotein A-I/ApoA1 Antibody (2083A) [Unconjugated]

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and Hep3B human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human Apolipoprotein A-I/ApoA1 Monoclonal Antibody (Catalog # MAB36641) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Apolipoprotein A-I/ApoA1 at approximately 25-30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HepG2 human hepatocellular carcinoma cell line was stained with Rabbit Anti-Human Apolipoprotein A-I/ApoA1 Monoclonal Antibody (Catalog # MAB36641, filled histogram) or isotype control antibody (Catalog # MAB1050, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Apolipoprotein A-I/ApoA1 was detected in immersion fixed human hepatocytes using Rabbit Anti-Human Apolipoprotein A-I/ApoA1 Monoclonal Antibody (Catalog # MAB36641) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell secretion. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Apolipoprotein A-I/ApoA1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit Anti-Human Apolipoprotein A-I/ApoA1 Monoclonal Antibody (Catalog # MAB36641) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Apolipoprotein A‑I/ApoA1 was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Rabbit Anti-Human Apolipoprotein A‑I/ApoA1 Monoclonal Antibody (Catalog # MAB36641) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Particle number increases proportionally with needle-to-processing time. (a) A set of four peripheral blood samples was each collected in n=4 biological replicates Streck Cell-Free DNA BCT. EV enrichment was performed by differential ultracentrifugation (dUC), followed by SEC. EV-enriched fractions were analyzed by NTA, BCA and WB. (b) Particle numbers obtained from the different needle-to-processing times, measured by NTA. Data normalized to the control condition of immediately processed blood. Mean of normalized value is displayed for each needle-to-processing time and statistical significance is assessed using one sample t test (Bonferroni-Holm-adjusted p-values) with *p=0.0175 (24 h), **p=0.0033 (72 h) and **p=0.0033 (1 week). (c) Median size of particles recovered in EV-enriched fractions from the different needle-to-processing times. Mean value is displayed for each needle-to-processing time and statistical significance is assessed using paired t test (Bonferroni-Holm-adjusted p-values) with **p=0.0014 (24 h vs. immediate processing), ***p=0.0006 (72 h vs. immediate processing) and ***p=0.0006 (1 week vs. immediate processing). In (b) and (c) symbols signify independent biological replicates. (d) Representative immunoblotting image to characterize particles obtained according to EV markers, cell-specific markers and apolipoprotein markers. Original blots are presented in Supplementary Fig. S10 online. (e)-(g) Quantification of band intensities in (d). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/40624148), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, Flow, ICC/IF
• Clone: 2083A
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

Apolipoprotein A-I/ApoA1 Antibody (2083A) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E. coli-derived recombinant human ApoA1
  Asp25-Gln267
  Accession # P02647
• Specificity: Detects human ApoA1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1-25 ug/mL
  • Immunohistochemistry 1-25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 0.2 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.
• Preservative: Sodium Azide
• 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 Apolipoprotein A-I/ApoA1 Antibody (2083A) [Unconjugated]

• Alp-1
• APOA1
• Apo-AI
• apoA-I
• Apolipoprotein A1
• Apolipoprotein AI
• Apolipoprotein A-I
• Brp-14
• Ltw-1
• Lvtw-1
• MGC117399
• Sep-1
• Sep-2

Background

ApoA1 is a major protein component of serum high-density lipoprotein (HDL) and is produced by the liver and small intestine. It is involved in reverse cholesterol transport from tissues to the liver. ApoA1 is synthesized as a 267 amino acid (aa) precursor from which a signal peptide and short propeptide are removed. Mature human ApoA1 shares 65% and 61% aa sequence identity with mouse and rat ApoA1, respectively.

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 Apolipoprotein A-I/ApoA1 Antibody (MAB36641)(1)

Publication using MAB36641

• Megan I. Mitchell, Iddo Z. Ben‐Dov, Christina Liu, Kenny Ye, Kar Chow, Yael Kramer et al. Extracellular Vesicle Capture by AnTibody of CHoice and Enzymatic Release (EV‐CATCHER): A customizable purification assay designed for small‐RNA biomarker identification and evaluation of circulating small‐EVs Journal of Extracellular Vesicles 2021-06-03 [PMID: 34122779] (Western Blot, Human)

Bioinformatics

• Uniprot:
  • P02647()