ADNP Antibody [Unconjugated]

Western blot shows lysates of embryonic rat cortical neuron/ glial cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse/Rat ADNP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5919) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for ADNP at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ADNP was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Goat Anti-Mouse/Rat ADNP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5919) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

ADNP was detected in perfusion fixed frozen sections of mouse brain (cerebellum) using Goat Anti-Mouse/Rat ADNP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5919) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001). Specific staining was localized to Purkinje cells and neurons in molecular cell layer. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

ADNP was detected in immersion fixed paraffin-embedded sections of mouse brain using Goat Anti-Mouse/Rat ADNP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5919) at 1.7 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004) or the HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Product Discontinued

ADNP Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse ADNP
  Leu941-Ala1102
  Accession # Q9H2P0
• Specificity: Detects mouse and rat ADNP in direct ELISAs and Western blots. In direct ELISAs, approximately 30% cross-reactivity with recombinant human ADNP is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ADNP
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 1-15 ug/mL
  • Western Blot 1 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. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for ADNP Antibody [Unconjugated]

• Activity-dependent neuroprotective protein
• activity-dependent neuroprotector homeobox
• activity-dependent neuroprotector
• ADNP homeobox 1
• ADNP
• ADNP1
• ADNP1KIAA0784activity-dependent neuroprotector homeobox protein

Background

ADNP (Activity-dependent neuroprotective protein) is an ADNF-related factor found in astrocytes and microvascular endothelial cells. Although its predicted MW is 124 kDa, it runs anomalously at 148‑150 kDa in SDS-PAGE. ADNP is inducible by VIP in astrocytes, and serves as a neuroprotector, both outside and inside the cell. Extracellularly, ADNP may protect synapse activity; intracellularly, ADNP interacts with tubulin, promoting microtubule assembly, and within the nucleus, it contributes to the SWI/SNF chromatin remodeling complex. Mouse ADNP is 1108 amino acids (aa) in length. It contains a neuroprotective peptide/NAP (aa 354‑361) embedded between nine C2H2-type Zn finger domains (aa 76‑685), a DNA-binding homeobox region (aa 752‑813) and a Leu-rich NLS that overlaps a protein internalization sequence (aa 787‑812). There are alternative start sites at Met229 and Met281. Over aa 947‑1108, mouse ADNP shares 98% and 93% aa identity with rat and human ADNP, 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 ADNP Antibody (AF5919)(3)

Publications using AF5919

• XiaoYun Sun, WenJun Yu, Li Li, YuHua Sun ADNP Controls Gene Expression Through Local Chromatin Architecture by Association With BRG1 and CHD4 Frontiers in Cell and Developmental Biology 2020-07-01 [PMID: 32714933]
• X Sun, X Peng, Y Cao, Y Zhou, Y Sun ADNP promotes neural differentiation by modulating Wnt/beta-catenin signaling Nat Commun, 2020-06-12;11(1):2984. 2020-06-12 [PMID: 32533114] (Co-IP, Western Blot, Mouse)
• Dresner E, Malishkevich A, Arviv C, Leibman Barak S, Alon S, Ofir R, Gothilf Y, Gozes I Novel evolutionary-conserved role for the activity-dependent neuroprotective protein (ADNP) family that is important for erythropoiesis. J Biol Chem, 2012-10-15;287(48):40173-85. 2012-10-15 [PMID: 23071114] (ChIP, Mouse)

Bioinformatics

• Gene Symbol: ADNP
• Uniprot:
  • Q9H2P0(Human)