Human Collagen Type IV, REH-P02462
Product Introduction
Human Collagen Type IV is a heterotrimeric protein composed of two α1-like and one α2-like gene products, representing the major structural element of basement membranes in multicellular tissues. Type IV collagen forms a sheet-like extracellular matrix that surrounds specific cell types, including adipocytes, skeletal muscle cells, cardiac muscle cells, and smooth muscle cells. It serves as a critical scaffold for basement membrane assembly and interacts with laminin and nidogen to establish the supramolecular architecture essential for tissue integrity, filtration, and cell-matrix signaling. Human Collagen Type IV is implicated in Alport syndrome and Goodpasture syndrome, as well as in severe rheumatic and dermatological diseases such as epidermolysis bullosa acquisita. This product is extracted from washed dissected human tissue into dilute acetic acid following mild pepsin treatment, and purified by differential salt precipitation. The resulting protein retains its native triple-helical conformation, with purity and structural integrity confirmed by SDS-PAGE and reactivity with collagen type-specific monoclonal antibodies.
Product Features
1. Native Triple-Helical Conformation: Extracted under mild conditions to preserve the native triple-helical structure of type IV collagen, essential for authentic biological activity.
2. High Purity: Purified by differential salt precipitation to >95% as determined by SDS-PAGE, ensuring reproducibility in experimental applications.
3. Basement Membrane Component: Provides the primary structural framework of basement membranes, facilitating studies of cell-matrix interactions, cell adhesion, and basement membrane biology.
4. Defined Binding Partners: Interacts specifically with laminin and nidogen, enabling reconstitution studies of basement membrane assembly.
5. Clinical Relevance: Implicated in autoimmune and genetic basement membrane disorders, providing a valuable tool for studying the molecular pathogenesis of Alport syndrome, Goodpasture syndrome, and autoimmune blistering diseases.
Specifications
Size: 1 mg
Catalog Number: REH-P02462
Protein: Human Collagen Type IV
Species: Human
Molecular Weight: ~300 kDa (heterotrimer)
Purity: >95%
Concentration: 0.5-5 mg/mL in 0.01 mol/L PBS
Form: Liquid purified protein
NCBI Accession: NM_001845.4
UniProt Accession: P02462
Binding Specificity: Laminin; Nidogen
Storage and Stability
Storage Conditions: Store at -20°C or -80°C. Avoid repeated freeze-thaw cycles as these may disrupt the native triple-helical conformation.
Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed. For working aliquots, store at 4°C and use within 1 week.
Protocol (For Reference Only)
Important: Collagen Type IV is a structural protein; handle gently to avoid shearing and maintain the native triple-helical conformation. Thaw on ice before use.
1. Coating of Cell Culture Surfaces: Dilute Human Collagen Type IV in sterile 0.01 mol/L PBS or sterile 0.05-0.1 M acetic acid to a working concentration of 5-10 µg/cm². Add sufficient volume to cover the culture surface and incubate at room temperature for 1-2 hours or at 37°C for 30-60 minutes. Aspirate the coating solution, rinse twice with sterile PBS, and allow to air dry in a sterile biosafety cabinet before seeding cells.
2. Collagen Gel Formation: For three-dimensional gel preparations, neutralize the acidic collagen solution with appropriate amounts of sterile 0.1 M NaOH and 10X concentrated culture medium or PBS on ice. Mix thoroughly to achieve a final neutral pH (7.2-7.4). Transfer to culture vessels and incubate at 37°C for 30-60 minutes to allow gel polymerization. Seed cells onto or within the collagen gel as required.
3. Cell Adhesion Assays: Coat 96-well plates with Human Collagen Type IV at concentrations ranging from 1-50 µg/mL in PBS overnight at 4°C. Block wells with 1% BSA for 1 hour at 37°C, seed cells at appropriate density, and incubate at 37°C for 1-2 hours. Quantify adherent cells by colorimetric or fluorometric methods.
4. Basement Membrane Reconstitution Studies: For in vitro basement membrane assembly assays, combine Human Collagen Type IV with purified laminin and nidogen in appropriate molar ratios. Incubate at 37°C and monitor network formation by electron microscopy or immunofluorescence.
5. Immunological Studies: Use as a substrate for ELISA or Western blot to screen for anti-collagen IV autoantibodies associated with Goodpasture syndrome, Alport syndrome, or epidermolysis bullosa acquisita. The optimal working dilution must be determined by the end user.
Precautions
1. Avoid repeated freeze-thaw cycles; aliquot into single-use volumes upon initial thawing and store at -20°C or -80°C.
2. The product is provided as a liquid in 0.01 mol/L PBS. Do not vortex; gently pipette to mix.
3. For coating applications, working concentrations must be optimized empirically for each cell type and experimental system.
4. The protein concentration range is 0.5-5 mg/mL; refer to the lot-specific certificate of analysis for the exact concentration of each batch.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the molecular composition of Human Collagen Type IV?
A1: Human Collagen Type IV is a heterotrimer composed of two α1-like chains and one α2-like chain. It forms a native triple-helical structure with a molecular weight of approximately 300 kDa.
Q2: What are the known binding partners of Collagen Type IV?
A2: Collagen Type IV specifically interacts with laminin and nidogen. These interactions are critical for the assembly and structural integrity of basement membranes.
Q3: What diseases are associated with Collagen Type IV?
A3: Collagen Type IV is implicated in Alport syndrome (a genetic disorder affecting basement membranes in the kidney, ear, and eye), Goodpasture syndrome (an autoimmune disease targeting the α3 chain of type IV collagen in kidney and lung basement membranes), and acquired autoimmune blistering diseases such as epidermolysis bullosa acquisita.
Q4: How should I store the product after opening?
A4: Aliquot into single-use volumes upon receipt and store at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Working aliquots may be stored at 4°C for up to 1 week.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of coating concentrations and assay conditions is strongly recommended for specific cell types and experimental systems.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: REH-P02462
Product Name: Human Collagen Type IV
Size: 1 mg
Price: CNY ¥3500.00 / USD $350.00 / EUR �420.00 / JPY ¥63000.00
2023 Version
