Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?

Name: Mouse TNF-alpha Quantikine HS ELISA Kit
Brand: R&D Systems
SKU: MHSTA50
Rating: 4.5 (2 reviews)

A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

Mouse TNF-alpha Quantikine HS ELISA Kit

Catalog #: MHSTA50
13 Citations 2 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MHSTA50

Control Products Available

QC228: Quantikine Immunoassay Control Set 913 for Mouse HS TNF-a

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Product Details

Citations (13)

FAQs

Supplemental Products

Reviews (2)

Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse TNF-alpha Quantikine HS ELISA Kit Summary

Assay Length

4.0 hours

Sample Type & Volume Required Per Well

Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)

Sensitivity

0.295 pg/mL

Assay Range

0.8 - 50 pg/mL (Serum, EDTA Plasma, Heparin Plasma)

Specificity

Natural and recombinant mouse TNF-alpha.

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Product Summary

The Quantikine HS Mouse TNF-alpha Immunoassay is a 4.0 hour solid phase ELISA designed to measure mouse TNF-alpha levels in serum and plasma. It contains E. coli-expressed recombinant mouse TNF-alpha and antibodies raised against the recombinant factor and has been shown to accurately quantitate recombinant mouse TNF-alpha. Results obtained using natural TNF-alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that the Quantikine HS Immunoassay kit can be used to determine relative mass values for natural mouse TNF-alpha.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Serum, EDTA Plasma, Heparin Plasma

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	6.63	15.1	29.2	6.29	15.2	28.5
Standard Deviation	0.257	0.695	1.49	0.552	1.18	1.94
CV%	3.9	4.6	5.1	8.8	7.8	6.8

Recovery

The recovery of mouse TNF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type	Average % Recovery	Range %
EDTA Plasma (n=4)	91	81-104
Heparin Plasma (n=4)	90	79-106
Serum (n=4)	86	77-99

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of mouse TNF-alpha were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

Scientific Data

N/A

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Mouse TNF-alpha ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF1A, is the prototypic ligand of the TNF superfamily (1). It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (2-5). TNF-alpha is also involved in a number of pathological conditions including asthma, Crohn's disease, rheumatoid arthritis, neuropathic pain, obesity, type 2 diabetes, septic shock, autoimmunity, and cancer (5-11).

Mouse TNF-alpha is synthesized as a 26 kDa type II transmembrane protein that consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (12). Within the ECD, mouse TNF-alpha shares 95% aa identity with rat, and 80% aa identity with canine, equine, feline, human, rabbit, and porcine TNF-alpha. It is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface (13). Cell surface TNF-alpha can both induce the lysis of tumor cells and virus infected cells, and generate its own downstream cell signaling following ligation by soluble TNF RI (14, 15). Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer containing the TNF-alpha extracellular domain (16-18).

TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (19, 20) and the hematopoietic cell-restricted 78-80 kDa TNF RII (21, 22), both of which are also expressed as homotrimers (1, 23). Both type I and type II receptors bind TNF-alpha with comparable affinity and can promote NFkB activation (24-27). Only TNF RI, however, contains a cytoplasmic death domain which triggers the activation of apoptosis (3, 28). Soluble forms of both types of receptors are released into human serum and urine, and can neutralize the biological activity of TNF (29-31).

Long Name:

Tumor Necrosis Factor alpha

Entrez Gene IDs:

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Alternate Names:

APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

Related Research Areas

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Mouse TNF-alpha Quantikine HS ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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Tumour extracellular vesicles and particles induce liver metabolic dysfunction
Authors: Wang, G;Li, J;Bojmar, L;Chen, H;Li, Z;Tobias, GC;Hu, M;Homan, EA;Lucotti, S;Zhao, F;Posada, V;Oxley, PR;Cioffi, M;Kim, HS;Wang, H;Lauritzen, P;Boudreau, N;Shi, Z;Burd, CE;Zippin, JH;Lo, JC;Pitt, GS;Hernandez, J;Zambirinis, CP;Hollingsworth, MA;Grandgenett, PM;Jain, M;Batra, SK;DiMaio, DJ;Grem, JL;Klute, KA;Trippett, TM;Egeblad, M;Paul, D;Bromberg, J;Kelsen, D;Rajasekhar, VK;Healey, JH;Matei, IR;Jarnagin, WR;Schwartz, RE;Zhang, H;Lyden, D;
Nature
Species: Mouse
Sample Types: Cell Culture Supernates, Plasma
Efficacy and safety of ketone ester infusion to prevent muscle weakness in a mouse model of sepsis-induced critical illness
Authors: R Weckx, C Goossens, S Derde, L Pauwels, S Vander Per, G Van den Be, L Langouche
Oncogene, 2022-06-22;12(1):10591.
Species: Mouse
Sample Types: Plasma
IL-6 Deficiency Attenuates Skeletal Muscle Atrophy by Inhibiting Mitochondrial ROS Production through the Upregulation of PGC-1alpha in Septic Mice
Authors: B Yang, X Yang, X Sun, J Shi, Y Shen, R Chen
Oxidative Medicine and Cellular Longevity, 2022-04-27;2022(0):9148246.
Species: Mouse
Sample Types: Cell Culture Supernates
Identification for antitumor effects of tramadol in a xenograft mouse model using orthotopic breast cancer cells
Authors: MH Kim, JR Lee, KJ Kim, JH Jun, HJ Hwang, W Lee, SH Nam, JE Oh, YC Yoo
Scientific Reports, 2021-11-11;11(1):22113.
Species: Mouse
Sample Types: Serum
Identification of the toxic threshold of 3-hydroxybutyrate-sodium supplementation in septic mice
Authors: R Weckx, C Goossens, S Derde, L Pauwels, S Vander Per, G Van den Be, L Langouche
Bmc Pharmacology & Toxicology, 2021-09-20;22(1):50.
Species: Mouse
Sample Types: Plasma
Scopolamine promotes neuroinflammation and delirium-like neuropsychiatric disorder in mice
Authors: SY Cheon, BN Koo, SY Kim, EH Kam, J Nam, EJ Kim
Scientific Reports, 2021-04-16;11(1):8376.
Species: Mouse
Sample Types: Tissue Homogenates
Orthopedic surgery-induced cognitive dysfunction is mediated by CX3CL1/R1 signaling
Authors: I Cho, JM Kim, EJ Kim, SY Kim, EH Kam, E Cheong, M Suh, BN Koo
Journal of Neuroinflammation, 2021-04-15;18(1):93.
Species: Mouse
Sample Types: Tissue Homogenates
Liver fibrosis-induced muscle atrophy is mediated by elevated levels of circulating TNF&alpha
Authors: T Kurosawa, M Goto, N Kaji, S Aikiyo, T Mihara, M Ikemoto-Ue, M Toyoda, N Kanazawa, T Nakazawa, M Hori, A Uezumi
Cell Death & Disease, 2021-01-07;12(1):11.
Species: Mouse
Sample Types: Serum
Modulation of Macrophage Polarization by Phospholipids on the Surface of Titanium
Authors: H Quan, Y Kim, L Wu, HC Park, HC Yang
Molecules, 2020-06-10;25(11):.
Species: Mouse
Sample Types: Cell Culture Supernates
CXCL14 Overexpression Attenuates Sepsis-Associated Acute Kidney Injury by Inhibiting Proinflammatory Cytokine Production
Authors: J Lv, ZL Wu, Z Gan, P Gui, SL Yao
Mediators Inflamm., 2020-03-31;2020(0):2431705.
Species: Mouse
Sample Types: Serum, Tissue Homogenates

Adipose tissue protects against sepsis-induced muscle weakness in mice: from lipolysis to ketones
Authors: C Goossens, R Weckx, S Derde, T Dufour, S Vander Per, L Pauwels, SE Thiessen, PP Van Veldho, G Van den Be, L Langouche
Crit Care, 2019-07-01;23(1):236.
Species: Mouse
Sample Types: Plasma

Microbubble-mediated delivery of human adenoviruses does not elicit innate and adaptive immunity response in an immunocompetent mouse model of prostate cancer
Authors: F De Carlo, L Thomas, B Brooke, ET Varney, R Nande, O Boskovic, GD Marshall, PP Claudio, CM Howard
J Transl Med, 2019-01-11;17(1):19.
Species: Mouse
Sample Types: Serum

Predominant gut Lactobacillus murinus strain mediates anti-inflammaging effects in calorie-restricted mice
Authors: F Pan, L Zhang, M Li, Y Hu, B Zeng, H Yuan, L Zhao, C Zhang
Microbiome, 2018-03-21;6(1):54.
Species: Mouse
Sample Types: Serum

FAQs

Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?
- A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

View all ELISA FAQs