GFR alpha-like Antibody [Unconjugated]

Western blot shows lysates of Neuro-2A mouse neuroblastoma cell line, mouse brain and mouse brain stem. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Mouse GFRa-like Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5728) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). Specific bands were detected for GFRa-like at approximately 25 and 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

GFR alpha ‑like was detected in immersion fixed paraffin-embedded sections of mouse spinal cord using Sheep Anti-Mouse GFR alpha ‑like Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5728) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with DAPI (blue). Specific staining was localized to dorsal and ventral horns. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

GFRa-like was detected in immersion fixed frozen sections of mouse brain using Sheep Anti-Mouse GFRa-like Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5728) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with Fluoro Nissl Green (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Neuronal activation in CPT-treated mice.DIO mice (weighing 40 to 42 g) received vehicle or CPT (1 mg kg−1); 60 min later, brains were harvested, and tissue sections were subjected to immunofluorescence staining with anti-c-Fos (red) and anti-GFRAL (green). (A) Representative immunofluorescence image of the AP/NTS in CPT- or vehicle-treated DIO mice showing the distribution of GFRAL neurons and the colocalization with the marker of neuronal activation c-Fos. (B) Quantification of CPT-induced c-Fos expression in the AP/NTS (n = 6 per group). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. The underlying data for this figure can be found in S1 Data. AP, area postrema; CPT, Camptothecin; DIO, diet-induced obese; GFRAL, GDNF receptor alpha-like; NTS, nucleus tractus solitarius. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35202387), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

GFR alpha-like Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant mouse GFR alpha -like isoform A
  Gln20-Gly349
  Accession # Q6SJE0
• Specificity: Detects mouse GFR alpha -like in direct ELISAs. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse (rm)GFR alpha -3 and rmGFR alpha -4 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: Gfral
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 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. 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 GFR alpha-like Antibody [Unconjugated]

• bA360D14.1
• C6orf144
• GDNF family receptor alpha-like
• GFR alpha like
• GFR alpha-like
• GFRAL
• GRAL
• IVFI9356
• UNQ9356

Background

GFR alpha -like (GDNF receptor-alpha-like) is a predicted 26 and 44 kDa glycoprotein (1) and distant member of the GDNFR family of proteins. It is expressed on both fetal and adult neurons of the CNS, and would appear to function as an anti-apoptotic molecule during neuronal stress. Mature mouse GFR alpha -like is a 374 amino acid (aa) type I transmembrane protein. It contains a 330 aa extracellular region that shows a Gas1 domain (aa 131‑210), and a 23 aa cytoplasmic tail. There is one splice variant that is likely soluble and shows a four aa substitution for aa 235‑393. Over aa 20‑349, mouse GFR alpha -like shares 72% aa identity with human GFR alpha -like.

1. Li, Z. et al. (2005) J. Neurochem. 95:361.

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 GFR alpha-like Antibody (AF5728)(4)

Publications using AF5728

• Beom Yong Lee, Jongwon Jeong, Inseong Jung, Hanchae Cho, Dokyung Jung, Jiwon Shin et al. GDNF family receptor alpha‐like antagonist antibody alleviates chemotherapy‐induced cachexia in melanoma‐bearing mice Journal of Cachexia, Sarcopenia and Muscle 2023-06-01 [PMID: 37017344]
• Beom Yong Lee, Jongwon Jeong, Inseong Jung, Hanchae Cho, Dokyung Jung, Jiwon Shin et al. GDNF family receptor alpha‐like antagonist antibody alleviates chemotherapy‐induced cachexia in melanoma‐bearing mice Journal of Cachexia, Sarcopenia and Muscle 2023-06-01 [PMID: 37017344] (Immunohistochemistry, Mouse)
• Ricardo J. Samms, Richard Cosgrove, Brandy M. Snider, Ellen C. Furber, Brian A. Droz, Daniel A. Briere et al. GIPR Agonism Inhibits PYY-Induced Nausea-Like Behavior Diabetes 2022-07-01 [PMID: 35499381]
• Chuchu Zhang, Judith A. Kaye, Zerong Cai, Yandan Wang, Sara L. Prescott, Stephen D. Liberles Area Postrema Cell Types that Mediate Nausea-Associated Behaviors Neuron 2021-02-01 [PMID: 33278342]

Bioinformatics

• Gene Symbol: Gfral
• Uniprot:
  • Q6SJE0(Rat)