Human Sequestosome 1 (SQSTM1) ELISA Kit
Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to SQSTM1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to SQSTM1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain SQSTM1, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of SQSTM1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
For Use with tissue homogenates, cell lysates or other biological fluids. For Research Use Only. Not for use in diagnostic procedures.
Target Information
Autophagy receptor required for selective macroautophagy (aggrephagy). Functions as a bridge between polyubiquitinated cargo and autophagosomes. Interacts directly with both the cargo to become degraded and an autophagy modifier of the MAP1 LC3 family (PubMed:16286508, PubMed:20168092, PubMed:24128730, PubMed:28404643, PubMed:22622177). Along with WDFY3, involved in the formation and autophagic degradation of cytoplasmic ubiquitin-containing inclusions (p62 bodies, ALIS/aggresome-like induced structures). Along with WDFY3, required to recruit ubiquitinated proteins to PML bodies in the nucleus (PubMed:24128730, PubMed:20168092). May regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD (By similarity). May be involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels. Involved in endosome organization by retaining vesicles in the perinuclear cloud: following ubiquitination by RNF26, attracts specific vesicle-associated adapters, forming a molecular bridge that restrains cognate vesicles in the perinuclear region and organizes the endosomal pathway for efficient cargo transport (PubMed:27368102). Promotes relocalization of 'Lys-63'-linked ubiquitinated STING1 to autophagosomes (PubMed:29496741). Acts as an activator of the NFE2L2/NRF2 pathway via interaction with KEAP1: interaction inactivates the BCR(KEAP1) complex, promoting nuclear accumulation of NFE2L2/NRF2 and subsequent expression of cytoprotective genes (PubMed:20452972, PubMed:28380357, PubMed:33393215).
| GENE ID | 8878 |
| SWISS PROT | Q13501 |
| SYNONYMS |
OSIL; PDB3; ZIP3; p62; p62B; p60; A170; Paget Disease Of Bone 3; Oxidative Stress Induced Like; EBI3-associated protein of 60 kDa; Ubiquitin-binding protein p62 |
Materials Supplied
| Kit Components | 96 Wells Quantity/Size |
|---|---|
| Pre-coated, ready-to-use 96-well strip plate | 1 plate |
| Plate sealer for 96 wells | 2 |
| Standard |
2 tubes |
| Diluent buffer | 1 bottle |
| Detection Reagent A | 1 bottle |
| Detection Reagent B | 1 bottle |
| TMB Substrate | 1 tube |
| Stop Solution | 1 tube |
| Wash Buffer (30 ℅ concentrate) | 1 tube |
| Product data sheet | 1 copy |
Storage
| Storage | The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C. |
Performance Characteristics
| REPEATABILITY |
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level SQSTM1 were tested 20 times on one plate, respectively. |
| SENSITIVITY | The minimum detectable dose was 0.125ng/mL. |
| ASSAY RANGE | 0.312-20ng/mL |
| SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of SQSTM1. No significant cross-reactivity or interference between SQSTM1 and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between SQSTM1 and all analogs, therefore, cross reactivity may still exist. |