SKU EK0375
Size 96wells/kit, with removable strips.
Reactivity Mouse
Sample Type cell culture supernates and serum.
Sensitivity <5pg/ml
Assay Range 31.2pg/ml-2000pg/ml

Overview

Product Name Mouse IFN Gamma PicoKine™ ELISA Kit
SKU/Catalog Number EK0375
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 Mouse IFN gamma. 96wells/kit, with removable strips.
Cite This Product Mouse IFN Gamma PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0375)
Sample Type cell culture supernates and serum..
Sensitivity <5pg/ml
Assay Range 31.2pg/ml-2000pg/ml
Immunogen Expression system for standard: E.coli; Immunogen sequence: H23-C155
Reactivity Mouse
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.

Assay Details

Kit Components

Catalog numberDescriptionQuantity
EK0375-CAP96-well plate precoated with anti-Mouse Ifng antibody1
EK0375-STlyophilized recombinant Mouse Ifng standard10ng/tube
EK0375-DAbiotinylated anti-Mouse Ifng antibody130ul(dilution 1:100)
AR1103Avidin-Biotin-Peroxidase Complex(ABC)130ul(dilution 1:100)
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4

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 Mouse IFN Gamma PicoKine™ ELISA Kit

(TMB reaction incubate at 37°C for 25-30min)

Concentration(pg/ml)02000200020002000200020002000
O.D.0.0490.1420.2530.3750.5951.0151.3551.997

Intra/Inter Assay Precision

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean(pg/ml)6822476070232686
Standard deviation2.9212.3233.443.0812.7630.18
CV(%)4.3%5.5%4.4%4.4%5.5%4.4%

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.

LotsLot1 (pg/ml)Lot2 (pg/ml)Lot3 (pg/ml)Lot4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 168696665671.582.3%
Sample 222420320824321915.627.1%
Sample 376083973188380360.687.5%
*number of samples for each test n=16.

*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 P01580
Gene Name Ifng
Protein Name Interferon gamma
Tissue Specificity Released primarily from activated T lymphocytes.
Alternative Names Interferon gamma;IFN-gamma;Ifng;
Subcellular Localization Secreted.
Molecular Weight 17907 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
Background Interferon-gamma(IFN-gamma) is an inflammatory cytokine that has been implicated in the development of fibrosis in inflamed tissues. The production of IFN-gamma, which is under genetic control, can influence the development of fibrosis in lung allografts. IFN-gamma is also produced by natural killer(NK) cells and most prominently by CD8 cytotoxic T cells, and is vital for the control of microbial pathogens. Interferon gamma is believed to be crucial for host defence against many infections. Genetically determined variability in IFN-gamma and expression might be important for the development of tuberculosis. IFN-gamma activates human macrophage oxidative metabolism and antimicrobial activity. In addition to having antiviral activity, IFN-gamma has important immunoregulatory functions. IFN-gamma plays an important role in the control of neointima proliferation.

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Publications

Protective immunity against acute toxoplasmosis in BALB/c mice induced by a DNA vaccine encoding?Toxoplasma gondii?elongation factor 1-alpha
3-(1H-Benzo[d]imidazol-6-yl)-5-(4-fluorophenyl)-1,2,4-oxadiazole (DDO7232), a Novel Potent Nrf2/ARE Inducer, Ameliorates DSS-Induced Murine Colitis and Protects NCM460 Cells against Oxidative Stress via ERK1/2 Phosphorylation
Immune Protection of Rhoptry Protein 21 (ROP21) of Toxoplasma gondii as a DNA Vaccine Against Toxoplasmosis
5-BDBD ameliorates an OVA-induced allergic asthma by the reduction of Th2 cytokines production
Herbal Components of a Novel Formula PSORI-CM02 Interdependently Suppress Allograft Rejection and Induce CD8+CD122+PD-1+ Regulatory T Cells
Silencing of suppressor of cytokine signaling 1 enhances the immunological effect of mucin 1-calreticulin-primed 4T1 cell-treated dendritic cells in breast cancer treatment
Establishment of a mouse model of lipopolysaccharide-induced neutrophilic nasal polyps
hsdS, Belonging to the Type I Restriction-Modification System, Contributes to the Streptococcus suis Serotype 2 Survival Ability in Phagocytes
The hemagglutinin-neuramidinase protein of Newcastle disease virus upregulates expression of the TRAIL gene in murine natural killer cells through the activation of Syk and NF-?B
The cryo-thermal therapy eradicated melanoma in mice by eliciting CD4+ T-cell-mediated antitumor memory immune response
Toxoplasma gondii Elongation Factor 1-Alpha (TgEF-1?) Is a Novel Vaccine Candidate Antigen against Toxoplasmosis
Genetically Engineered Virus Nanofibers as an Efficient Vaccine for Preventing Fungal Infection
A deleterious role for Th9/IL-9 in hepatic fibrogenesis
Immunological adjuvant effect of the peptide fraction from the larvae of Musca domestica
Stimulation of dendritic cells by DAMPs in ALA-PDT treated SCC tumor cells
Immunostimulative Activity of Low Molecular Weight Chitosans in RAW264.7 Macrophages
Changes of cytokine levels in a mouse model of post-infectious irritable bowel syndrome
3-(3-Pyridylmethylidene)-2-indolinone Reduces the Severity of Colonic Injury in a Murine Model of Experimental Colitis
Minocycline treatment ameliorates interferon-alpha- induced neurogenic defects and depression-like behaviors in mice
Interleukin-22 ameliorates liver fibrogenesis by attenuating hepatic stellate cell activation and downregulating the levels of inflammatory cytokines
Virus-Like Particles of Chimeric Recombinant Porcine Circovirus Type 2 as Antigen Vehicle Carrying Foreign Epitopes
DNA vaccination with a gene encoding?Toxoplasma gondii?Deoxyribose Phosphate Aldolase (TgDPA) induces partial protective immunity against lethal challenge in mice
Structure and Antitumor and Immunomodulatory Activities of a Water-Soluble Polysaccharide fromDimocarpus longan?Pulp
Exogenous interleukin-10 alleviates allergic inflammation but inhibits local interleukin-10 expression in a mouse allergic rhinitis model
Construction and Immunogenicity of DNA Vaccines Encoding Fusion Protein of Porcine IFN-?1 and GP5 Gene of Porcine Reproductive and Respiratory Syndrome Virus
Hybrid phage displaying SLAQVKYTSASSI induces protection against?Candida albicans?challenge in BALB/c mice
Immunomodulatory Activity and Protective Effects of Polysaccharide from Eupatorium adenophorum Leaf Extract on Highly Pathogenic H5N1 Influenza Infection
Dynamically expressed microRNA-15b modulates the activities of CD8+?T lymphocytes in mice with Lewis lung carcinoma
A novel DNA vaccine expressing the Ag85A-HA2 fusion protein provides protection against influenza A virus and?Staphylococcus aureus
Molecular Adjuvant Ag85A Enhances Protection against Influenza A Virus in Mice Following DNA Vaccination
Xu B, Zhang P, Li W, Liu R, Tang J, Fan H. Front Microbiol. 2017 Aug 9;8:1524. doi: 10.3389/fmicb.2017.01524. eCollection 2017. hsdS, Belonging to the Type I Restriction-Modification System, Contributes to the Streptococcus suis Serotype 2 Survival Ability in Phagocytes
Qin S, Gao Z, Liu Y, Liu C, Wang J, Zou LL. Oncol Lett. 2018 Feb;15(2):1630-1638. doi: 10.3892/ol.2017.7477. Epub 2017 Nov 23. Silencing of suppressor of cytokine signaling 1 enhances the immunological effect of mucin 1-calreticulin-primed 4T1 cell-treated dendritic cells in breast cancer treatment
Wang S1, Zhang H1, Xi Z1, Huang J1, Nie J1, Zhou B1, Deng Y2, Tao Z2. Exp Ther Med. 2017 Dec;14(6):5275-5282. doi: 10.3892/etm.2017.5208. Epub 2017 Sep 27. Establishment of a mouse model of lipopolysaccharide-induced neutrophilic nasal polyps
Lu C, Liu H, Jin X, Chen Y, Liang CL, Qiu F, Dai Z. Front Pharmacol. 2018 Feb 12;9:88. doi: 10.3389/fphar.2018.00088. eCollection 2018. Herbal Components of a Novel Formula PSORI-CM02 Interdependently Suppress Allograft Rejection and Induce CD8+CD122+PD-1+ Regulatory T Cells
Jiang, Z., Chi, J., Han, B., & Liu, W. (2017). Preparation and pharmacological evaluation of norcantharidin-conjugated carboxymethyl chitosan in mice bearing hepatocellular carcinoma. Carbohydrate Polymers, 174, 282-290. Advance online publication. doi: 10.1016/j.carbpol.2017.06.072
Liu, C., Huang, R., Yao, R., & Yang, A. (2017). The Immunotherapeutic Role of Bacterial Lysates in a Mouse Model of Asthma. Lung, 1-7. Advance online publication. doi: 10.1007/s00408-017-0003-8.
He K, Liu P, Xu LX. Cell Death Dis. 2017 Mar 23;8(3):e2703. doi: 10.1038/cddis.2017.125.The cryo-thermal therapy eradicated melanoma in mice by eliciting CD4+ T-cell-mediated antitumor memory immune response.
Zhong G, Cheng X, Long H, He L, Qi W, Xiang T, Zhao Z, Zhu B. J Transl Med. 2013 Mar 21;11:71. Doi: 10.1186/1479-5876-11-71. Dynamically Expressed Microrna-15B Modulates The Activities Of Cd8+ T Lymphocytes In Mice With Lewis Lung Carcinoma.
Sofian M, Aghakhani A, Farazi Aa, Banifazl M, Eslamifar A, Rashidi N, Khadem Sadegh A, Ramezani A. Hepat Mon. 2012 Dec;12(12):E6156. Doi: 10.5812/Hepatmon.6156. Epub 2012 Dec 29. Serum Profile Of T Helper 1 And T Helper 2 Cytokines In Hepatitis C ...
Pan Zy, Wang H. Int Immunopharmacol. 2014 May;20(1):229-37. Doi: 10.1016/J.Intimp.2014.03.004. Epub 2014 Mar 19. Synergistic Interaction Between Choline And Aspirin Against Acute Inflammation Induced By Carrageenan And Lipopolysaccharide.

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
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
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: 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: 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 mouse ifn gamma elisa kit, mouse ifi elisa kit, mouse ifng elisa kit, mouse interferon gamma elisa kit