INPP5F Antibody, Biotin conjugated

Introduction

The INPP5F Antibody, Biotin conjugated is a specialized immunological reagent designed for detecting the INPP5F protein, a phosphoinositide 4-phosphatase involved in cellular membrane trafficking and phosphoinositide metabolism. This antibody is conjugated with biotin, enabling its use in high-sensitivity assays like enzyme-linked immunosorbent assays (ELISA) and immunoprecipitation via streptavidin-based detection systems.

Key Features

• Target Protein: INPP5F (Sac2), a 4-phosphatase that regulates phosphatidylinositol 4-phosphate (PI4P) levels in early endosomes .

• Conjugation: Biotin allows for versatile detection using streptavidin-horseradish peroxidase (HRP) or fluorescent streptavidin in downstream assays.

• Applications: Validated for ELISA, Western blot (WB), and immunofluorescence (IF) in human and murine samples .

3.1. Western Blotting

• Detects INPP5F in lysates of cells with endosomal defects (e.g., Rab5-positive compartments) .

• Example: In studies of hepatocellular carcinoma (HCC), this antibody confirmed INPP5F overexpression in tumor tissues (TCGA-LIHC cohort) .

3.2. Immunofluorescence

• Localizes INPP5F to early endosomes (co-staining with Rab5/EEA1 markers) .

• Application: Visualizing INPP5F translocation in HCC cells (cytoplasmic vs. nuclear) .

3.3. ELISA

• Quantifies INPP5F levels in biological fluids or lysates .

• Use Case: Monitoring INPP5F expression in cerebrospinal fluid (CSF) for neurodegenerative studies .

4.1. Endocytosis Studies

• Sac2/INPP5F interacts with OCRL in Rab5-positive endosomes to regulate PI(4,5)P₂ metabolism .

• Knockout models (Sac2 KO MEFs) exhibit impaired transferrin internalization, highlighting its role in clathrin-mediated endocytosis .

4.2. Cancer Research

• Overexpression in HCC correlates with poor prognosis and promotes aerobic glycolysis via Notch signaling .

• Mechanism: INPP5F interacts with ASPH to activate c-MYC and cyclin E1, enhancing tumor growth .

4.3. Neuroregeneration

• Sac2 deletion in mice enhances axon regeneration post-spinal cord injury (SCI) by modulating membrane trafficking .

• Safety Profile: No adverse effects in INPP5F-null mice, unlike PTEN inhibitors .

Validation and Quality Control

• Western Blot: Detects a ~65 kDa band in HEK293 lysates .

• Immunofluorescence: Colocalizes with Rab5 (Pearson’s R > 0.7) in COS7 cells .

• ELISA: Linear range: 1–100 ng/mL (inter-assay CV < 10%) .