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Catalog No: OKCD04072 (Formerly GWB-U3NO5F)
Size:96WELLS
Price: $625.00
SKU
OKCD04072
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Datasheets/ManualsPrintable datasheet for HMGB1 ELISA Kit (Mouse) (OKCD04072)
COA DatasheetClickHereto download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityMouse|Mus musculus
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit Linearity
Matrix1:21:41:81:16
Serum (n=5)99-105%78-92%87-95%94-102%
EDTA Plasma (n=5)82-90%83-97%80-88%85-96%
Heparin Plasma(n=5)97-105%86-104%86-102%95-105%
ELISA Kit PrincipleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to High Mobility Group Protein 1 (HMGB1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Mobility Group Protein 1 (HMGB1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain High Mobility Group Protein 1 (HMGB1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of High Mobility Group Protein 1 (HMGB1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range46.88-3,000pg/mL
ELISA Kit Recovery
MatrixRecovery Range (%)Average (%)
Serum (n=5)86-9592
EDTA Plasma (n=5)90-104101
Heparin Plasma(n=5)80-9489
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Mobility Group Protein 1 (HMGB1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Mobility Group Protein 1 (HMGB1) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
HMGB1 Microplate96 Wells (12 x 8 Well strips)
HMGB1 Lyophilized Standard2 x 3 ng
100X Biotinylated HMGB1 Detector Antibody1 x 120 uL
100X Avidin-HRP Conjugate1 x 120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
Stop Solution1 x 6 mL
TMB Substrate1 x 9 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeSerum, plasma and other biological fluids
Sensitivity18.29 pg/mL
Publications

HMGB1 promotes ductular reaction and tumorigenesis in autophagy-deficient livers. J Clin Invest. 128, 2419-2435 (2018) 29558368

Loss of High-Mobility Group Box 1 (HMGB1) Protein in Rods Accelerates Rod Photoreceptor Degeneration After Retinal Detachment. Invest Ophthalmol Vis Sci. 61, 50 (2020). 32460314

Streptococcal sagA activates a proinflammatory response in mast cells by a sublytic mechanism. Cell Microbiol. 21, e13064 (2019). 31155820

Assay InfoAssay Methodology: Quantitative Sandwich ELISA
Protocol Information
Description
Gene SymbolGm21596|Hmgb1
Gene Full Namehigh mobility group box 1|predicted gene, 21596
Alias Symbolsamph;amphoterin;high mobility group protein 1;high mobility group protein B1;Hmg1;HMG-1;p3;p30;SBP;SBP-1.
NCBI Gene Id100862258|15289
Protein NameHigh mobility group protein B1
Description of TargetMultifunctional redox sensitive protein with various roles in different cellular compartments. In the nucleus is one of the major chromatin-associated non-histone proteins and acts as a DNA chaperone involved in replication, transcription, chromatin remodeling, V(D)J recombination, DNA repair and genome stability. Proposed to be an universal biosensor for nucleic acids. Promotes host inflammatory response to sterile and infectious signals and is involved in the coordination and integration of innate and adaptive immune responses. In the cytoplasm functions as sensor and/or chaperone for immunogenic nucleic acids implicating the activation of TLR9-mediated immune responses, and mediates autophagy. Acts as danger associated molecular pattern (DAMP) molecule that amplifies immune responses during tissue injury. Released to the extracellular environment can bind DNA, nucleosomes, IL-1 beta, CXCL12, AGER isoform 2/sRAGE, lipopolysaccharide (LPS) and lipoteichoic acid (LTA), and activates cells through engagement of multiple surface receptors. In the extracellular compartment fully reduced HMGB1 (released by necrosis) acts as a chemokine, disulfide HMGB1 (actively secreted) as a cytokine, and sulfonyl HMGB1 (released from apoptotic cells) promotes immunological tolerance (PubMed:23519706, PubMed:23446148, PubMed:23994764, PubMed:25048472). Has proangiogenic activity (PubMed:16365390). May be involved in platelet activation. Binds to phosphatidylserine and phosphatidylethanolamide. Bound to RAGE mediates signaling for neuronal outgrowth. May play a role in accumulation of expanded polyglutamine (polyQ) proteins (By similarity).
Uniprot IDP63158
Protein Accession #NP_065390
Nucleotide Accession #NM_000079
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