Detection of Mouse/Rat ADNP by Western Blot.
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 in RAW 264.7 Mouse Cell Line.
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 in Mouse Brain.
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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
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.
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