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| SKU | EK0514 |
|---|---|
| Size | 96wells/kit, with removable strips. |
| Reactivity | Rat |
| Sample Type | cell culture supernates, serum, plasma(EDTA) and urine. |
| Sensitivity | <1pg/ml |
| Assay Range | 15.6pg/ml-1000pg/ml |
Overview
| Product Name | Rat TGF Beta 1 PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0514 |
| Storage & Handling | Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.) |
| Size | 96wells/kit, with removable strips. |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of activated Rat TGF beta 1. 96wells/kit, with removable strips. |
| Cite This Product | Rat TGF Beta 1 PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0514) |
| Sample Type | cell culture supernates, serum, plasma(EDTA) and urine..
Anticoagulant(s): EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants thata are not specified above or contact us to check for feasibility. |
| Sensitivity | <1pg/ml |
| Assay Range | 15.6pg/ml-1000pg/ml |
| Immunogen | Expression system for standard: CHO; Immunogen sequence: A279-S390 |
| Reactivity | Rat |
| Cross Reactivity | There is cross-reactivity with TGFbeta2, TGFbeta3, TGFbeta5<1%. |
Assay Details
Kit Components
| Catalog number | Description | Quantity |
| EK0514-CAP | 96-well plate precoated with anti-Rat Tgfb1 antibody | 1 |
| EK0514-ST | lyophilized recombinant Rat Tgfb1 standard | 10ng/tube |
| EK0514-DA | biotinylated anti-Rat Tgfb1 antibody | 130ul(dilution 1:100) |
| AR1103 | Avidin-Biotin-Peroxidase Complex(ABC) | 130ul(dilution 1:100) |
| AR1106-1 | sample diluent buffer | 30ml |
| AR1106-2 | antibody diluent buffer | 12ml |
| AR1106-3 | ABC diluent buffer | 12ml |
| AR1104 | TMB color developing agent | 10ml |
| AR1105 | TMB stop solution | 10ml |
| PLA-SEA | Adhesive cover | 4 |
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Rat TGF Beta 1 PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-20min)
| Concentration(pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.072 | 0.157 | 0.218 | 0.346 | 0.646 | 1.088 | 1.638 | 2.230 |
Intra/Inter Assay Precision
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean(pg/ml) | 27 | 161 | 431 | 28 | 156 | 422 |
| Standard deviation | 1.59 | 9.82 | 23.7 | 1.73 | 9.98 | 26.16 |
| CV(%) | 5.5%% | 6.1% | 5.5% | 6.2% | 6.4% | 6.2% |
Reproducibility
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
Reproducibility
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
| Lots | Lot1 (pg/ml) | Lot2 (pg/ml) | Lot3 (pg/ml) | Lot4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 27 | 31 | 29 | 32 | 29 | 1.92 | 6.6% |
| Sample 2 | 161 | 143 | 162 | 145 | 152 | 8.78 | 5.7% |
| Sample 3 | 431 | 413 | 424 | 455 | 430 | 15.4 | 3.5% |
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Target Info
Protein Target Info (Source: Uniprot.org)
| Uniprot Id | P17246 |
|---|---|
| Gene Name | Tgfb1 |
| Protein Name | Transforming growth factor beta-1 |
| Tissue Specificity | Abundant in the bone matrix. . |
| Alternative Names | Transforming growth factor beta-1;TGF-beta-1;Latency-associated peptide;LAP;Tgfb1; |
| Subcellular Localization | Secreted, extracellular space, extracellular matrix . |
| Molecular Weight | 44329 MW |
*if product is indicated to react with multiple species, protein info is based on the human gene.
Ontology
| Protein Function | 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. Can promote either T- helper 17 cells (Th17) or regulatory T-cells (Treg) lineage differentiation in a concentration-dependent manner. At high concentrations, leads to FOXP3-mediated suppression of RORC and down-regulation of IL-17 expression, favoring Treg cell development. 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). . |
|---|---|
| Background | Transforming growth factor-beta1(TGF-beta1) is a multifunctional peptide that controls proliferation, differentiation, and other functions in many cell types. Many cells synthesize TGF-beta and essentially all of them have specific receptors for this peptide. TGF-beta regulates the actions of many other peptide growth factors and determines a positive or negative direction of their effects. TGFbeta1 is known for its potent and diverse biological effects, including immune regulation, and cell growth and differentiation. TGFbeta1 is also an important mediator of bone remodeling. TGFbeta1, a potent keratinocyte growth inhibitor, has been shown to be overexpressed in keratinocytes in certain inflammatory skin diseases and has been thought to counteract the effects of other growth factors at the site of inflammation. TGF-beta1, a multifunctional cytokine with fibrogenic properties, has been implicated in the pathogenesis of the vascular and target organ complications of hypertension. TGF-beta1 may also regulate blood pressure via stimulation of endothelin-1 and/or renin secretion. TGFbeta1 is secreted as a latent form, which consists of its mature form and a latency-associated peptide(beta1-LAP) in either the presence or the absence of additional latent TGF-beta1-binding protein. The standard product used in this kit is recombinant TGFbeta1 with the molecular mass of 25KDa. |
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Publications
Theaflavin alleviates inflammatory response and brain injury induced by cerebral hemorrhage via inhibiting the nuclear transcription factor kappa ?-related pathway in rats
Hydroethanolic Extract of Strychnos pseudoquina Accelerates Skin Wound Healing by Modulating the Oxidative Status and Microstructural Reorganization of Scar Tissue in Experimental Type I Diabetes
Danggui buxue tang suppresses high glucose-induced proliferation and extracellular matrix accumulation of mesangial cells via inhibiting lncRNA PVT1
Efficacies of rosiglitazone and retinoin on bleomycin-induced pulmonary fibrosis in rats
Nitric Oxide Synthase Activity Correlates with OGG1 in Ozone-Induced Lung Injury Animal Models
Bacillus Calmette-Guerin alleviates airway inflammation and remodeling by preventing TGF-?1 induced epithelial?mesenchymal transition
A New Oleanolic Acid Derivative against CCl4-Induced Hepatic Fibrosis in Rats
LSKL peptide alleviates subarachnoid fibrosis and hydrocephalus by inhibiting TSP1-mediated TGF-?1 signaling activity following subarachnoid hemorrhage in rats
3,4-dihydroxyphenylethanol suppresses irradiation-induced pulmonary fibrosis in adult rats
Resveratrol Ameliorates Diabetes-induced Renal Damage through Regulating the Expression of TGF-?1, Collagen IV and Th17/Treg-related Cytokines in Rats
The green tea extract epigallocatechin-3-gallate inhibits irradiation-induced pulmonary fibrosis in adult rats
Evening primrose oil ameliorates platelet aggregation and improves cardiac recovery in myocardial-infarct hypercholesterolemic rats
Correlation between apoptosis and TGF-?1 expression in the mucosal epithelium of rat small intestine in a cold stress state
Over-production of nitric oxide by oxidative stress-induced activation of the TGF-?1/PI3K/Akt pathway in mesangial cells cultured in high glucose
Transforming growth factor ?1-induced astrocyte migration is mediated in part by activating 5-lipoxygenase and cysteinyl leukotriene receptor 1
The Ayurvedic Medicine?Salacia oblonga?Attenuates Diabetic Renal Fibrosis in Rats: Suppression of Angiotensin II/AT1 Signaling
Emodin protects rat liver from CCl4-induced fibrogenesis via inhibition of hepatic stellate cells activation
Effect of Oxymatrine on the TGFbeta-Smad signaling pathway in rats with CCl4-induced hepatic fibrosis
The Ayurvedic Medicine
Emodin protects rat liver from CCl4-induced fibrogenesis via inhibition of hepatic stellate cells activation
Effect of Oxymatrine on the TGFbeta-Smad signaling pathway in rats with CCl4-induced hepatic fibrosis
Sarandy MM, Miranda LL, Alto
Zhang S, Li J, Li Y, Liu Y, Guo H, Xu X. Front Physiol. 2017 Apr 27;8:249. doi: 10.3389/fphys.2017.00249. eCollection 2017. Nitric Oxide Synthase Activity Correlates with OGG1 in Ozone-Induced Lung Injury Animal Models
Luo, H., Liu, B., Zhao, L., He, J., Li, T., Zha, L.,..., & Yu, Z. (2017). Galectin-3 mediates pulmonary vascular remodeling in hypoxia-induced pulmonary arterial hypertension. Journal of the American Society of Hypertension. Advance online publication. doi: 10.1016/j.jash.2017.07.009
Hou Sx, Zhu Wj, Pang Mq, Jeffry J, Zhou Ll. Food Chem Toxicol. 2014 Feb;64:57-64. Doi: 10.1016/J.Fct.2013.11.022. Epub 2013 Nov 26. Protective Effect Of Iridoid Glycosides From Paederia Scandens (Lour.) Merrill (Rubiaceae) On Uric Acid Nephropathy...
Sharifi N, Amani R, Hajiani E, Cheraghian B. Endocrine. 2014 Sep;47(1):70-80. Doi: 10.1007/S12020-014-0336-5. Epub 2014 Jun 27. Does Vitamin D Improve Liver Enzymes, Oxidative Stress, And Inflammatory Biomarkers In Adults With Non-Alcoholic Fatty ...
Gao L, Huang W, Li J. Mol Cell Biochem. 2013 Oct;382(1-2):185-91. Doi: 10.1007/S11010-013-1733-4. Epub 2013 Jun 26. Nox1 Abet Mesangial Fibrogenesis Via Inos Induction In Diabetes.
Customer Q&As
Q: Does the TMB color developing agent contain H2O2?
A: Yes, the TMB color developing agent contain H2O2.
Q: Does this ELISA kit contain any product produced in humans or primates?
Keywords: component, ingredient
Keywords: component, ingredient
A: None of the components in our ELISA kits are produced in humans or primates.
Q: What is the volume of the recombinant protein control? What is its shelf life?
Keywords: expiration, storage, temperature, size
Keywords: expiration, storage, temperature, size
A: The volume of the control is around 100pg. If unopened, the shelf life of the control is the same as the whole kit - "Store at 4˚C for 6 months, at -20˚C for 12 months." The reconstituted control can be stored at -20˚C for 2 days.
Q: Will it be okay to run the experiment if I accidentally used the wrong buffer (e.g. sample diluent buffer) for antibody dilution instead of the antibody diluent buffer? Keywords: dilution, replace, substitute, sample buffer
A: Using the sample diluent buffer for antibody dilution instead of the antibody diluent buffer will negatively affect the test result to some extent. The values of both standard and sample might be lower than the normal values.
Q: In the "Preparation" section of the protocol, step 2 says to "activate the sample". What is the difference between active form TGF beta 1 and inactive form TGF beta 1 in analysis?
Keywords: sample activation, manual, data calculation, experimental result
A: TGFb in a biological sample will generally be present in one of two forms, either as LAP+TGFb (=latent TGFb) or free TGFb. The ELISA only recognizes free TGFb. Therefore, if you do not activate your sample, you will only be measuring the free TGFb in the sample. If you run the assay twice, one without activation (=free TGFb) and once with activation (=free TGFb + latent TGFb), the difference in TGFb values from the two assays will be the amount of latent TGFb in the sample.
Q: The protocol says to use neutral PBS and provides a recipe. Could I use neutral PBS made with KCl or KH2PO4 (common constituents for most PBS recipes) instead?
Keywords: protocol, alternative buffer, applications, reagent recipe
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
Q: What is your recommended protocol on preparing tissues (such as bone) for ELISA analysis?
Keyword: sample preparation, treatment procedures
A: The protocol for bone tissue is as follows -
Bone Tissue: Extract demineralized bone samples in 4 M Guanidine-HCl and protease inhibitors. Dissolve the final sample in 2 M Guanidine-HCl.
However, we did not test tissue samples with the Elisa kits so a pilot experiment is recommended.
Q: Why are my O.D. values different than your values on the datasheet?
Keyword: troubleshooting, reference
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
Q: On the ELISA kit datasheet it says "HRP substrate TMB was used to visualize HRP enzymatic reaction." What is HRP and which part of the kit contains HRP?
Keyword: kit components, reagents, protocol
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
Q: Do you offer any ELISA kits that can work with whole blood samples instead of plasma or urine?
Keyword: applications
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
Q: Does silicic acid (formula SiOH4) affect the results of the ELISA assay?
Keyword: protocol, reagents
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
Q: Is there lot-to-lot variation of the ELISA kits? What are Boster's general services when I have questions about your kits?
Keyword: technical support, help, customer service
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
Q: The results of my standards do not look correct, what could be the problem?
Keyword: troubleshoot, protocol
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
Q: What is the well depth of the 96 well plates for the ELISA kits?
Keyword: well capacity, product size
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
Q: The kit does not include wash solution, what should I do?
Keyword: wash buffer
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
Q: For your ELISA kits, are the capture and detection antibodies polyclonals or monoclonals?
Keyword: antibody clonality
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
Q: Do your ELISA kits come with sealants or plate covers?
Keyword: storage, sequential use
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
Q: What is the concentration of Sodium Azide in your Elisa kits?
Keyword: preservative
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
Q: Are there positive and negative controls available for my ELISA kit?
Keyword: positive control, negative control
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
Q: Why do I get positive results for my knockout (KO) model when used as a control?
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
Q: How long do I soak my plate in the wash buffer?
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
Q: Can I use Tween in my wash buffer?
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
Q: What plate type do I use to set up the microplate reader?
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the normal level of this protein in my sample of interest?
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
Q: Is the plate separable? Can I use only part of the kit and save the rest for later? How many samples can I run with one kit?
A: Yes the plate is separable. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
Q: Does the kit contain sample preparation reagents? How do I prepare my samples?
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
Q: Can this ELISA kit work on brain tissue homogenate, cell culture supernatant, saliva, urine, serum, whole blood or any other sample type?
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
Q: Can this ELISA kit react with the pro-form of the target protein? Can this ELISA kit react with an isoform of the protein?
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our techincal support.
Q: Can this ELISA kit react with human, mouse, rat or other species?
A: If the kit is reactive to another commonly used species (human, mouse, and/or rat), we would have listed it as a separate product. If you want to check cross-reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ identical, there is a high chance of cross-reactivity for your species of interest.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to rat tgfb elisa kit, rat tgfb1 elisa kit, rat tgfbeta elisa kit, rat tgf-beta-1 elisa kit