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HMGB1 ELISA Kit (Rat) (OKCD04073)

96 Wells
$579.00
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Gene Symbol:
HMGB1
Official Gene Full Name:
high mobility group box 1
NCBI Gene Id:
25459
Alias Symbols:
Ac2-008, Amphoterin, Heparin-binding protein p30, High mobility group protein 1, High mobility group protein B1, Hmg1, Hmg-1, HMG-1, MGC93598, MGC93599
Description of Target:
Multifunctional 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.
Protein Name:
High mobility group protein B1
Swissprot Id:
P63159
Protein Accession #:
NP_037095
Nucleotide Accession #:
NM_012963
Predicted Species Reactivity:
Rat
Sample Type:
Serum, plasma and other biological fluids
Sensitivity:
6.7 pg/mL
Kit Range:
15.6-1,000 pg/mL
Kit Reproducibility:
Three samples concentrations were measured in replicate within an assay plate and across replicate assays to assess Intra- and Mean Inter-Assay Precision.
Mean Intra-Assay Precision - <10% (n = 20)
Mean Inter-Assay Precision - <12% (n = 8)
Kit Duration:
~3 Hours
Kit Principle:
Aviva Systems Biology HMGB1 ELISA Kit (Rat) (OKCD04073) is based on standard sandwich enzyme-linked immuno-sorbent assay technology. An antibody specific for HMGB1 has been pre-coated onto a 96-well plate (12 x 8 Well Strips). Standards or test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for HMGB1 is added, incubated and followed by washing. Avidin-Peroxidase Conjugate is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is produced through the addition of TMB substrate which is catalyzed by HRP generating a blue color product that changes to yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm is quantitatively proportional to the amount of sample HMGB1 captured in the well.
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
Kit Linearity:
Matrix1:21:41:81:16
Serum (n=5)78-98%79-93%96-104%80-105%
EDTA Plasma (n=5)91-98%85-92%84-101%88-99%
Heparin Plasma(n=5)88-102%98-105%79-92%96-105%
Kit Recovery:
MatrixRecovery Range (%)Average (%)
Serum (n=5)83-10391
EDTA Plasma (n=5)96-103101
Heparin Plasma(n=5)79-9987
Kit Detection Method:
Colorimetric, OD450 nm
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express HMGB1.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express HMGB1.
Datasheets/Manuals:
Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Reconstitution and Storage:
Store as indicated in product manual.
Assay Info:
Assay Methodology: Quantitative Sandwich ELISA

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128/05/2018 19:39
  • Quality:
  • Overall Experience:
“The kit instructions were easy to follow”

Submitted by:

Emory University

Claudia Espinosa-Garcia

HMGB1

  1. Sample: Rat serum
  2. Sample preparation: Serum was previously prepared and frozen at -80C. For 1:100 fold dilution, I added 10 uL of test sample to 990 uL of Standard Diluent as suggested.    
  3. Were these samples validated or characterized as positive and/or negative controls? HMGB1 is a protein ubiquitously expressed in the nuclei of mammalian cells. There are several reports measuring HMGB1 levels in rat serum, plasma, CSF samples (Kim et al., J Neurosci. 2006; 26(24):6413-21; Lee et al., Mol Brain. 2016; 9(1):81; Wang et al., Sci Rep. 2017; 7:46243;  Kim et al., Cell Death Dis. 2018; 9(4):426). Thus, it was expected to detect HMGB1 in my samples. 
  4. What dilution worked for your samples? From my previous experience with ELISA, I decided to use undiluted serum and 1:100 fold dilution. I observed that dilution 1:100 resulted in higher HMGB1 levels than undiluted serum.
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