| Description |
The Human High mobility group protein B1 (HMGB1) ELISA Kit allows for the in vitro quantitative determination of HMGB1 concentrations in serum, plasma, tissue homogenates, or cell lysates. This kit exclusively recognizes human HMGB1. HMGB1 is a nonhistone nucleoprotein and also an extracellular inflammatory cytokine. Extracellular HMGB1 acting as a damage-associated molecular pattern (DAMP) binds with its pattern recognition receptors (PRRs), mediating the proliferation, differentiation, mobilization, and senescence of hematopoietic stem cells (HSCs). HMGB1 also participates in the formation of the inflammatory bone marrow (BM) microenvironment by activating proinflammatory signaling pathways. It plays an important role in the pathogenesis of hematopoietic malignancies. Extensively DNA-bound intracellular HMGB1 is involved in transcriptional modulation, DNA replication and repair, telomere maintenance, and nucleosome assembly. The detection mechanism of this kit is based on the Sandwic... Read more |
| Target Name |
high-mobility group box 1 |
| Alternative Names |
Amphoterin ELISA Kit; Chromosomal protein; nonhistone; HMG1 ELISA Kit; DKFZp686A04236 ELISA Kit; High mobility group 1 ELISA Kit; High mobility group box 1 ELISA Kit; High mobility group protein 1 ELISA Kit; High mobility group protein B1 ELISA Kit; high-mobility group (nonhistone chromosomal) protein 1 ELISA Kit; HMG-1 ELISA Kit; HMG1 ELISA Kit; HMG3 ELISA Kit; HMGB 1 ELISA Kit; HMGB1 ELISA Kit; HMGB1_HUMAN ELISA Kit; NONHISTONE CHROMOSOMAL PROTEIN HMG1 ELISA Kit; SBP 1 ELISA Kit; Sulfoglucuronyl carbohydrate binding protein ELISA Kit |
| Abbreviation |
HMGB1 |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma, tissue homogenates, cell lysates |
| Detection Range |
78 pg/mL-5000 pg/mL |
| Sensitivity |
19.5 pg/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Epigenetics and Nuclear Signaling |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of human HMGB-1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
90 |
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| Range % |
84-99 |
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| 1:2 |
Average % |
88 |
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| Range % |
82-94 |
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| 1:4 |
Average % |
95 |
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| Range % |
89-102 |
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| 1:8 |
Average % |
94 |
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| Range % |
88-100 |
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| Recovery |
| The recovery of human HMGB-1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
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| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
98 |
90-102 |
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| EDTA plasma (n=4) |
91 |
85-100 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
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| pg/ml |
OD1 |
OD2 |
Average |
Corrected |
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| 5000 |
1.912 |
1.831 |
1.872 |
1.780 |
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| 2500 |
1.454 |
1.369 |
1.412 |
1.320 |
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| 1250 |
0.839 |
0.789 |
0.814 |
0.722 |
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| 625 |
0.456 |
0.487 |
0.472 |
0.380 |
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| 312 |
0.283 |
0.297 |
0.290 |
0.198 |
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| 156 |
0.199 |
0.202 |
0.201 |
0.109 |
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| 78 |
0.103 |
0.108 |
0.106 |
0.014 |
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| 0 |
0.089 |
0.094 |
0.092 |
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| Materials provided |
- A micro ELISA plate --- The 96-well plate has been pre-coated with anti-human HMGB1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard --- Dilute a bottle of standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled HMGB1 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator that can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
