HMGB1 ELISA Kit (Mouse)

• Reproducibility:

  Intra-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%

• Gene Name:

  High mobility group box 1|predicted gene, 21596

• Gene Aliases:

  Amph; amphoterin; high mobility group protein 1; high mobility group protein B1; Hmg1; HMG-1; p3; p30; SBP; SBP-1.

• Gene ID:

  100862258|15289

• Accession Number:

  NP_065390

• Reactivity:

  Mouse|Mus musculus

• 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. 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) .

• Type:

  ELISA Kit

• Applications:

  Enzyme-linked Immunosorbent assay-Sandwich

• Detection Method:

  Colorimetric

• Assay Principle:

  The 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.

• Assay Protocol:

  Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

• Sample Type:

  Serum, plasma and other biological fluids

• Detection Range:

  46.88-3,000pg/mL

• Recovery:

  Matrix Recovery Range (%) Average (%) _x005F Serum (n=5) 86-95 92 _x005F EDTA Plasma (n=5) 90-104 101 _x005F Heparin Plasma (n=5) 80-94 89 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 86-95 92 Serum (n=5) 86-95 92 EDTA Plasma (n=5) 90-104 101 EDTA Plasma (n=5) 90-104 101 Heparin Plasma (n=5) 80-94 89 Heparin Plasma (n=5) 80-94 89

• Sensitivity:

  18.29 pg/mL

• Reconstitution:

  2°C to 8°C|-20°C

• Components:

  Component Amount _x005F HMGB1 Microplate 96 Wells (12 x 8 Well strips) _x005F HMGB1 Lyophilized Standard 2 x 3 ng _x005F 100X Biotinylated HMGB1 Detector Antibody 1 x 120 uL _x005F 100X Avidin-HRP Conjugate 1 x 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F Stop Solution 1 x 6 mL _x005F TMB Substrate 1 x 9 mL _x005F Component Amount Component Amount HMGB1 Microplate 96 Wells (12 x 8 Well strips) HMGB1 Microplate 96 Wells (12 x 8 Well strips) HMGB1 Lyophilized Standard 2 x 3 ng HMGB1 Lyophilized Standard 2 x 3 ng 100X Biotinylated HMGB1 Detector Antibody 1 x 120 uL 100X Biotinylated HMGB1 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

• Notes:

  Formerly GWB-U3NO5F

• NCBI Gene Symbol:

  Gm21596|Hmgb1

• Protein Name:

  High mobility group protein B1

• Gene Name URL:

  Gm21596|Hmgb1

• CAS Number:

  7732-18-5

• Nucleotide Accession Number:

  NM_000079