Description
Rat Transforming growth factor beta-1 (Tgfb1) ELISA Kit
The Rat Transforming Growth Factor Beta 1 (TGF-β1) ELISA Kit is a highly reliable assay designed for the precise measurement of TGF-β1 levels in rat serum, plasma, and cell culture supernatants. This kit provides exceptional sensitivity and specificity, ensuring accurate and consistent results for a variety of research applications.TGF-β1 is a key regulatory cytokine involved in various cellular processes, including cell growth, differentiation, and immune response.
Dysregulation of TGF-β1 has been linked to numerous diseases, such as fibrosis, cancer, and autoimmune disorders, making it a valuable target for therapeutic interventions and biomarker studies.With its superior performance and ease of use, the Rat TGF-β1 ELISA Kit is an essential tool for researchers seeking to investigate the role of TGF-β1 in disease pathology and potential treatment strategies. Order yours today and streamline your TGF-β1 research efforts.
| Product Name: | Rat Transforming growth factor beta-1 (Tgfb1) ELISA Kit |
| SKU: | RTEB0097 |
| Size: | 96T |
| Target: | Rat Transforming growth factor beta-1 (Tgfb1) |
| Synonyms: | Transforming growth factor beta-1, TGF-beta-1, Tgfb1 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Rat |
| Detection Range: | 15.6-1000pg/mL |
| Sensitivity: | 7.82pg/mL |
| Intra CV: | 5.2% | ||||||||||||||||||||
| Inter CV: | 7.4% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Transforming growth factor beta-1: Multifunctional protein that regulates the growth and differentiation of various cell types and is involved in various processes, such as normal development, immune function, microglia function and responses to neurodegeneration (By similarity). Activation into mature form follows different steps: following cleavage of the proprotein in the Golgi apparatus, Latency-associated peptide (LAP) and Transforming growth factor beta-1 (TGF-beta-1) chains remain non-covalently linked rendering TGF-beta-1 inactive during storage in extracellular matrix (By similarity). At the same time, LAP chain interacts with 'milieu molecules', such as LTBP1, LRRC32/GARP and LRRC33/NRROS that control activation of TGF-beta-1 and maintain it in a latent state during storage in extracellular milieus. TGF-beta-1 is released from LAP by integrins (ITGAV:ITGB6 or ITGAV:ITGB8): integrin-binding to LAP stabilizes an alternative conformation of the LAP bowtie tail and results in distortion of the LAP chain and subsequent release of the active TGF-beta-1 (By similarity). Once activated following release of LAP, TGF-beta-1 acts by binding to TGF-beta receptors (TGFBR1 and TGFBR2), which transduce signal (By similarity). While expressed by many cells types, TGF-beta-1 only has a very localized range of action within cell environment thanks to fine regulation of its activation by Latency-associated peptide chain (LAP) and 'milieu molecules' (PubMed:2070682). Plays an important role in bone remodeling: acts as a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts (PubMed:2070682). Can promote either T-helper 17 cells (Th17) or regulatory T-cells (Treg) lineage differentiation in a concentration-dependent manner (By similarity). At high concentrations, leads to FOXP3-mediated suppression of RORC and down-regulation of IL-17 expression, favoring Treg cell development (By similarity). At low concentrations in concert with IL-6 and IL-21, leads to expression of the IL-17 and IL-23 receptors, favoring differentiation to Th17 cells (By similarity). Stimulates sustained production of collagen through the activation of CREB3L1 by regulated intramembrane proteolysis (RIP). Mediates SMAD2/3 activation by inducing its phosphorylation and subsequent translocation to the nucleus. Can induce epithelial-to-mesenchymal transition (EMT) and cell migration in various cell types (By similarity). |
| Uniprot: | P17246 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant rat Transforming growth factor beta-1 proprotein |
| Sub Unit: | Homodimer; disulfide-linked (By similarity). Interacts with the serine proteases, HTRA1 and HTRA3: the interaction with either inhibits TGFB1-mediated signaling. The HTRA protease activity is required for this inhibition. May interact with THSD4; this interaction may lead to sequestration by FBN1 microfibril assembly and attenuation of TGFB signaling (By similarity). Interacts with CD109, DPT and ASPN. Latency-associated peptide: Homodimer; disulfide-linked. Latency-associated peptide: Interacts with Transforming growth factor beta-1 (TGF-beta-1) chain; interaction is non-covalent and maintains (TGF-beta-1) in a latent state; each Latency-associated peptide (LAP) monomer interacts with TGF-beta-1 in the other monomer. Latency-associated peptide: Interacts with LTBP1; leading to regulate activation of TGF-beta-1. Latency-associated peptide: Interacts with LRRC32/GARP; leading to regulate activation of TGF-beta-1 on the surface of activated regulatory T-cells (Tregs). Interacts with LRRC33/NRROS; leading to regulate activation of TGF-beta-1 in macrophages and microglia. Latency-associated peptide: Interacts (via cell attachment site) with integrins ITGAV and ITGB6 (ITGAV:ITGB6), leading to release of the active TGF-beta-1 (By similarity). Latency-associated peptide: Interacts with NREP; the interaction results in a decrease in TGFB1 autoinduction (By similarity). Latency-associated peptide: Interacts with HSP90AB1; inhibits latent TGFB1 activation. Transforming growth factor beta-1: Homodimer; disulfide-linked. Transforming growth factor beta-1: Interacts with TGF-beta receptors (TGFBR1 and TGFBR2), leading to signal transduction (By similarity). |
| Research Area: | Cancer |
| Subcellular Location: | Transforming growth factor beta-1 Secreted |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | TGFB1: Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. Many cells synthesize TGFB1 and have specific receptors for it. It positively and negatively regulates many other growth factors. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts. Homodimer; disulfide-linked, or heterodimer with TGFB2. Secreted and stored as a biologically inactive form in the extracellular matrix in a 290 kDa complex (large latent TGF-beta1 complex) containing the TGFB1 homodimer, the latency-associated peptide (LAP), and the latent TGFB1 binding protein-1 (LTBP1). The complex without LTBP1 is known as the'small latent TGF-beta1 complex'. Dissociation of the TGFB1 from LAP is required for growth factor activation and biological activity. Release of the large latent TGF-beta1 complex from the extracellular matrix is carried out by the matrix metalloproteinase MMP3. May interact with THSD4; this interaction may lead to sequestration by FBN1 microfibril assembly and attenuation of TGFB signaling. Interacts with the serine proteases, HTRA1 and HTRA3: the interaction with either inhibits TGFB1-mediated signaling. The HTRA protease activity is required for this inhibition. Interacts with CD109, DPT and ASPN. Activated in vitro at pH below 3.5 and over 12.5. Highly expressed in bone. Abundantly expressed in articular cartilage and chondrocytes and is increased in osteoarthritis (OA). Co-localizes with ASPN in chondrocytes within OA lesions of articular cartilage. Belongs to the TGF-beta family. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Secreted; Secreted, signal peptide Cellular Component: axon; cell soma; cell surface; cytoplasm; extracellular matrix; extracellular space; microvillus; nucleus; proteinaceous extracellular matrix; secretory granule Molecular Function:antigen binding; cytokine activity; enzyme binding; glycoprotein binding; protein heterodimerization activity; protein homodimerization activity; protein N-terminus binding; protein serine/threonine kinase activator activity; punt binding; transforming growth factor beta receptor binding Biological Process: activation of NF-kappaB transcription factor; adaptive immune response based on somatic recombination of immune receptors built from immunoglobulin superfamily domains; aging; ATP biosynthetic process; cell activation; cell cycle arrest; cell migration; cell proliferation; cellular calcium ion homeostasis; chondrocyte differentiation; common-partner SMAD protein phosphorylation; defense response to fungus, incompatible interaction; endoderm development; epidermal growth factor receptor signaling pathway; epithelial to mesenchymal transition; evasion of host defenses by virus; female pregnancy; germ cell migration; gut development; heart development; hemopoietic progenitor cell differentiation; hyaluronan catabolic process; inflammatory response; inner ear development; intercellular junction assembly and maintenance; lipopolysaccharide-mediated signaling pathway; lung development; lymph node development; mammary gland development; MAPKKK cascade; membrane protein intracellular domain proteolysis; mitotic cell cycle checkpoint; mononuclear cell proliferation; morphogenesis of a branching structure; myelination; myeloid dendritic cell differentiation; negative regulation of blood vessel endothelial cell migration; negative regulation of cell cycle; negative regulation of cell differentiation; negative regulation of cell growth; negative regulation of cell proliferation; negative regulation of cell-cell adhesion; negative regulation of DNA replication; negative regulation of epithelial cell proliferation; negative regulation of fat cell differentiation; negative regulation of immune response; negative regulation of interleukin-17 production; negative regulation of mitotic cell cycle; negative regulation of myoblast differentiation; negative regulation of neuroblast proliferation; negative regulation of ossification; negative regulation of phagocytosis; negative regulation of protein amino acid phosphorylation; negative regulation of release of sequestered calcium ion into cytosol; negative regulation of skeletal muscle development; negative regulation of T cell activation; negative regulation of T cell proliferation; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; neural tube closure; neural tube development; Notch signaling pathway; oligodendrocyte development; organ regeneration; phosphate metabolic process; positive regulation of apoptosis; positive regulation of blood vessel endothelial cell migration; positive regulation of bone mineralization; positive regulation of cell migration; positive regulation of cell proliferation; positive regulation of cellular protein metabolic process; positive regulation of chemotaxis; positive regulation of collagen biosynthetic process; positive regulation of epithelial cell proliferation; positive regulation of exit from mitosis; positive regulation of fibroblast proliferation; positive regulation of histone acetylation; positive regulation of histone deacetylation; positive regulation of interleukin-17 production; positive regulation of isotype switching to IgA isotypes; positive regulation of MAP kinase activity; positive regulation of odontogenesis; positive regulation of peptidyl-serine phosphorylation; positive regulation of peptidyl-tyrosine phosphorylation; positive regulation of phosphoinositide 3-kinase activity; positive regulation of protein amino acid dephosphorylation; positive regulation of protein amino acid phosphorylation; positive regulation of protein complex assembly; positive regulation of protein import into nucleus; positive regulation of protein kinase B signaling cascade; positive regulation of protein secretion; positive regulation of regulatory T cell differentiation; positive regulation of smooth muscle cell differentiation; positive regulation of superoxide release; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; positive regulation of vascular permeability; protein amino acid phosphorylation; protein export from nucleus; protein import into nucleus, translocation; protein kinase B signaling cascade; receptor catabolic process; regulation of binding; regulation of cell growth; regulation of cell proliferation; regulation of DNA binding; regulation of gene expression; regulation of interleukin-23 production; regulation of MAPKKK cascade; regulation of protein import into nucleus; regulation of regulatory T cell differentiation; regulation of sodium ion transport; regulation of striated muscle development; regulation of transforming growth factor beta receptor signaling pathway; regulatory T cell differentiation; response to drug; response to estradiol stimulus; response to glucose stimulus; response to hypoxia; response to organic cyclic substance; response to organic substance; response to progesterone stimulus; response to radiation; response to vitamin D; response to wounding; salivary gland morphogenesis; SMAD protein complex assembly; SMAD protein nuclear translocation; T cell activation; T cell differentiation; T cell homeostasis; tolerance induction to self antigen; transforming growth factor beta receptor signaling pathway; ureteric bud development; vasculogenesis; ventricular cardiac muscle morphogenesis; wound healing |
| NCBI Summary: | binds the TGFbeta receptor; plays a role in regulation of cell growth and proliferation; induces synthesis of extracellular matrix proteins and may play a role in fibrosis [RGD, Feb 2006] |
| UniProt Code: | P17246 |
| NCBI GenInfo Identifier: | 135677 |
| NCBI Gene ID: | 59086 |
| NCBI Accession: | P17246.1 |
| UniProt Secondary Accession: | P17246,Q53YM8, |
| UniProt Related Accession: | P17246 |
| Molecular Weight: | 44,329 Da |
| NCBI Full Name: | Transforming growth factor beta-1 |
| NCBI Synonym Full Names: | transforming growth factor, beta 1 |
| NCBI Official Symbol: | Tgfb1 |
| NCBI Official Synonym Symbols: | Tgfb |
| NCBI Protein Information: | transforming growth factor beta-1; TGF-beta-1; transforming growth factor, beta-1 |
| UniProt Protein Name: | Transforming growth factor beta-1 |
| UniProt Synonym Protein Names: | |
| Protein Family: | Transforming growth factor |
| UniProt Gene Name: | Tgfb1 |
| UniProt Entry Name: | TGFB1_RAT |
| 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. |
