Description
Human E-selectin (SELE) ELISA Kit
The Human E-Selectin (SELE) ELISA Kit is a highly reliable and accurate tool designed for the precise measurement of E-selectin levels in human samples including serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring consistent and reproducible results for a wide range of research applications.E-selectin is a critical cell adhesion molecule that plays a key role in inflammation and immune response. Elevated levels of E-selectin have been associated with various inflammatory conditions, making it a valuable biomarker for studying diseases such as cardiovascular disorders, autoimmune diseases, and inflammatory conditions.
By using the Human E-Selectin ELISA Kit, researchers can accurately assess E-selectin levels in different biological samples, allowing for a better understanding of the role of E-selectin in various diseases and potentially aiding in the development of targeted therapies. This kit provides a valuable tool for advancing research in the fields of inflammation, immunity, and disease pathogenesis.
| Product Name: | Human E-selectin (SELE) ELISA Kit |
| SKU: | HUEB0108 |
| Size: | 96T |
| Target: | Human E-selectin (SELE) |
| Synonyms: | CD62 antigen-like family member E, Endothelial leukocyte adhesion molecule 1, Leukocyte-endothelial cell adhesion molecule 2, ELAM-1, LECAM2, CD62E, ELAM1 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 0.156-10ng/mL |
| Sensitivity: | 0.039ng/mL |
| Intra CV: | 3.9% | ||||||||||||||||||||
| Inter CV: | 7.1% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Cell-surface glycoprotein having a role in immunoadhesion. Mediates in the adhesion of blood neutrophils in cytokine-activated endothelium through interaction with SELPLG/PSGL1. May have a role in capillary morphogenesis. |
| Uniprot: | P16581 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human E-selectin |
| Sub Unit: | Interacts with SELPLG/PSGL1 and PODXL2 through the sialyl Lewis X epitope. SELPLG sulfation appears not to be required for this interaction. |
| Research Area: | Immunology |
| Subcellular Location: | Cell membrane Single-pass type I membrane protein |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | SELE: Cell-surface glycoprotein having a role in immunoadhesion. Mediates in the adhesion of blood neutrophils in cytokine-activated endothelium through interaction with PSGL1/SELPLG. May have a role in capillary morphogenesis. Belongs to the selectin/LECAM family. |
| UniProt Protein Details: | Protein type:Membrane protein, integral Chromosomal Location of Human Ortholog: 1q22-q25 Cellular Component: cortical cytoskeleton; extracellular space; perinuclear region of cytoplasm; plasma membrane; integral to membrane; coated pit; caveola; lipid raft Molecular Function:protein binding; transmembrane receptor activity; phospholipase binding; sialic acid binding Biological Process: positive regulation of leukocyte migration; positive regulation of receptor internalization; actin filament-based process; calcium-mediated signaling; response to lipopolysaccharide; leukocyte migration during inflammatory response; heterophilic cell adhesion; leukocyte adhesion; phospholipase C activation; regulation of inflammatory response; blood coagulation; inflammatory response; leukocyte tethering or rolling; leukocyte migration Disease: Hypertension, Essential |
| NCBI Summary: | The protein encoded by this gene is found in cytokine-stimulated endothelial cells and is thought to be responsible for the accumulation of blood leukocytes at sites of inflammation by mediating the adhesion of cells to the vascular lining. It exhibits structural features such as the presence of lectin- and EGF-like domains followed by short consensus repeat (SCR) domains that contain 6 conserved cysteine residues. These proteins are part of the selectin family of cell adhesion molecules. Adhesion molecules participate in the interaction between leukocytes and the endothelium and appear to be involved in the pathogenesis of atherosclerosis. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P16581 |
| NCBI GenInfo Identifier: | 126180 |
| NCBI Gene ID: | 6401 |
| NCBI Accession: | P16581.1 |
| UniProt Secondary Accession: | P16581,P16111, A2RRD6, |
| UniProt Related Accession: | P16581 |
| Molecular Weight: | 66,655 Da |
| NCBI Full Name: | E-selectin |
| NCBI Synonym Full Names: | selectin E |
| NCBI Official Symbol: | SELE |
| NCBI Official Synonym Symbols: | ELAM; ESEL; CD62E; ELAM1; LECAM2 |
| NCBI Protein Information: | E-selectin; ELAM-1; endothelial adhesion molecule 1; CD62 antigen-like family member E; endothelial leukocyte adhesion molecule 1; leukocyte endothelial cell adhesion molecule 2; leukocyte-endothelial cell adhesion molecule 2 |
| UniProt Protein Name: | E-selectin |
| UniProt Synonym Protein Names: | CD62 antigen-like family member E; Endothelial leukocyte adhesion molecule 1; ELAM-1; Leukocyte-endothelial cell adhesion molecule 2; LECAM2; CD_antigen: CD62E |
| Protein Family: | E-selectin |
| UniProt Gene Name: | SELE |
| UniProt Entry Name: | LYAM2_HUMAN |
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10mL | -20°C |
| Assay Diluent B | 10mL | -20°C |
| Detection Reagent A | 120µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Wash Buffer | 30mL | 4°C |
| Substrate | 10mL | 4°C |
| Stop Solution | 10mL | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
| Step | |
| 1. | Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C. |
| 2. | Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
