Human SOCS1 ELISA Kit
- SKU:
- HUFI01288
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O15524
- Sensitivity:
- 0.094ng/ml
- Range:
- 0.156-10ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- SOCS1, Suppressor of cytokine signaling 1, SOCS-1, Tec-interacting protein 3, TIP-3, JAK-binding protein, JAB, STAT-induced STAT inhibitor 1, SSI-1, SSI1, TIP3, CIS1, CISH1, JABTec-interacting protein 3, JAK binding protein, SOCS-1CIS1, SSI-1CISH1, S
- Reactivity:
- Human
- Research Area:
- Developmental Biology
Frequently bought together:
Description
Human SOCS1 ELISA Kit
STAT-induced STAT inhibitor (SSI), also known as suppressor of cytokine signalling (SOCS), is a gene that encodes a member of the STAT-induced STAT inhibitor (SSI) family. Cytokine-inducible negative regulators of cytokine signalling are members of the SSI family. A subset of cytokines, including IL2, IL3, erythropoietin, CSF2/GM-CSF, and interferon (IFN)-gamma, can increase the production of SOCS1. SOCS1 codes for a protein that works downstream of cytokine receptors and participates in a negative feedback loop that reduces cytokine signalling. The involvement of SOCS1 as a regulator of IFN-gamma action, which is necessary for proper postnatal growth and survival, was shown by knockout studies in mice. Autoinflammatory Syndrome, Familial, and Lipoprotein Quantitative Trait Locus are all diseases linked to SOCS1.
| Product Name: | Human SOCS1 ELISA Kit |
| Product Code: | HUFI01288 |
| Size: | 96 Assays |
| Alias: | SOCS1, Suppressor of cytokine signaling 1, SOCS-1, Tec-interacting protein 3, TIP-3, JAK-binding protein, JAB, STAT-induced STAT inhibitor 1, SSI-1, SSI1, TIP3, CIS1, CISH1, JABTec-interacting protein 3, JAK binding protein, SOCS-1CIS1, SSI-1CISH1, SSI1cytokine-inducible SH2 protein 1, STAT induced SH3 protein 1, STAT-induced STAT inhibitor 1 |
| Detection method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human SOCS1 concentrations in serum plasma and other biological fluids. |
| Sensitivity: | 0.094ng/ml |
| Range: | 0.156-10ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Human SOCS1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human SOCS1 in samples. | ||||||||||||||||
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human SOCS1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. | ||||||||||||||||
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| CV(%): | Intra-Assay: CV<8% Inter-Assay: CV<10% |
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/-20°C |
| Sample/Standard Dilution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody(Concentrated) | 120ul | 4°C (Protect from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate(SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protect from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer(25X) | 30ml | 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
| Uniprot | O15524 |
| UniProt Protein Function: | SOCS1: SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction. SOCS1 is involved in negative regulation of cytokines that signal through the JAK/STAT3 pathway. Through binding to JAKs, inhibits their kinase activity. In vitro, also suppresses Tec protein- tyrosine activity. Appears to be a major regulator of signaling by interleukin 6 (IL6) and leukemia inhibitory factor (LIF). Regulates interferon-gamma mediated sensory neuron survival. Probable substrate recognition component of an ECS (Elongin BC-CUL2/5-SOCS-box protein) E3 ubiquitin ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Seems to recognize JAK2. SOCS1 appears to be a negative regulator in IGF1R signaling pathway. Interacts with multiple activated signaling proteins of the tyrosine kinase signaling pathway including JAK family kinases, TEC, KIT, GRB2 and VAV. Binding to JAKs is mediated through the KIR and SH2 domains to a phosphorylated tyrosine residue within the JAK JH1 domain. Binds the SH3 domain of GRB2 via diproline determinants in the N-terminus, and the N-terminal regulatory domain of VAV. Interacts with the Elongin BC complex (TCEB1 and TCEB2). Component of an ECS CBC(SOCS1) E3 ubiquitin-protein ligase complex which contains Elongin BC, CUL5, RBX1 and SOCS1. Interacts (via SH2 domain and SOCS box) with TRIM8. Interacts with AXL, CUL2 and FGFR3. Interacts with INSR. By a subset of cytokines including those belonging to the interferon, interleukin and colony-stimulating factor families. Expressed in all tissues with high expression in spleen, small intestine and peripheral blood leukocytes. |
| UniProt Protein Details: | Protein type:Inhibitor Chromosomal Location of Human Ortholog: 16p13.13 Cellular Component: cytoplasm; cytoplasmic membrane-bound vesicle; cytosol; nucleus Molecular Function:insulin-like growth factor receptor binding; protein binding; protein kinase inhibitor activity; kinase inhibitor activity; protein kinase binding Biological Process: fat cell differentiation; regulation of protein amino acid phosphorylation; cytokine and chemokine mediated signaling pathway; regulation of growth; negative regulation of tyrosine phosphorylation of Stat3 protein; negative regulation of insulin receptor signaling pathway; protein ubiquitination; negative regulation of protein kinase activity; JAK-STAT cascade; negative regulation of JAK-STAT cascade |
| NCBI Summary: | This gene encodes a member of the STAT-induced STAT inhibitor (SSI), also known as suppressor of cytokine signaling (SOCS), family. SSI family members are cytokine-inducible negative regulators of cytokine signaling. The expression of this gene can be induced by a subset of cytokines, including IL2, IL3 erythropoietin (EPO), CSF2/GM-CSF, and interferon (IFN)-gamma. The protein encoded by this gene functions downstream of cytokine receptors, and takes part in a negative feedback loop to attenuate cytokine signaling. Knockout studies in mice suggested the role of this gene as a modulator of IFN-gamma action, which is required for normal postnatal growth and survival. [provided by RefSeq, Jul 2008] |
| UniProt Code: | O15524 |
| NCBI GenInfo Identifier: | 20178097 |
| NCBI Gene ID: | 8651 |
| NCBI Accession: | O15524.1 |
| UniProt Secondary Accession: | O15524,O15097, Q9NSA7, |
| UniProt Related Accession: | O15524 |
| Molecular Weight: | |
| NCBI Full Name: | Suppressor of cytokine signaling 1 |
| NCBI Synonym Full Names: | suppressor of cytokine signaling 1 |
| NCBI Official Symbol: | SOCS1 |
| NCBI Official Synonym Symbols: | JAB; CIS1; SSI1; TIP3; CISH1; SSI-1; SOCS-1 |
| NCBI Protein Information: | suppressor of cytokine signaling 1; TIP-3; JAK binding protein; JAK-binding protein; Tec-interacting protein 3; STAT induced SH3 protein 1; STAT-induced STAT inhibitor 1; cytokine-inducible SH2 protein 1 |
| UniProt Protein Name: | Suppressor of cytokine signaling 1 |
| UniProt Synonym Protein Names: | JAK-binding protein; JAB; STAT-induced STAT inhibitor 1; SSI-1; Tec-interacting protein 3; TIP-3 |
| Protein Family: | Suppressor of cytokine signaling |
| UniProt Gene Name: | SOCS1 |
| UniProt Entry Name: | SOCS1_HUMAN |
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Protocol |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
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. |
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