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Rat HMGB-1(High Mobility Group Protein B1) ELISA Kit

Citations(19)

  • Cat.No.:E-EL-R0505

  • Reactivity: Rat
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Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609
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Product Details

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 31.25-2000 pg/mL
Sensitivity 18.75 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Rat HMGB-1 in samples. No significant cross-reactivity or interference between Rat HMGB-1 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Rat HMGB-1 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat HMGB-1. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat HMGB-1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat HMGB-1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat HMGB-1. You can calculate the concentration of Rat HMGB-1 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips 		-20℃, 12 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL 		-20℃(Protect from light), 12 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 12 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat HMGB-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat HMGB-1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 92.38 254.88 974.98 84.22 250.98 977.51
Standard deviation 5.35 10.81 32.47 5.51 10.39 30.01
CV (%) 5.79 4.24 3.33 6.54 4.14 3.07

Recovery

The recovery of Rat HMGB-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 87-103 94
EDTA plasma (n=8) 93-106 98
Cell culture media (n=8) 91-104 97

Linearity

Samples were spiked with high concentrations of Rat HMGB-1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

SynonymsHMGB1, HMG1, HMG3, SBP-1, Amphoterin
Research AreaCancer, Epigenetics and Nuclear Signaling

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
elisa assay procedure 3

3. Aspirate and wash the plate for 3 times
elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 6

6. Add 50μL Stop Solution
elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. MEDIATORS OF INFLAMMATION (2020) IF: 4.711
    Exploring the Mechanism on the Medullary Visceral Zone Inhibiting the Cholinergic Anti-inflammatory Pathway Induced by Sepsis

    DOI: 10.1155/2020/1320278

    PMID: 33061821

    		 Sample: Serum
  2. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 6.208
    The Role of IGL-2 Preservation Solution on Rat Livers during SCS and HOPE

    DOI: 10.3390/ijms232012615

    PMID: 36293465

    		 Sample: Livers
  3. Intensive Care Medicine Experimental (2019)
    Use of a high platelet-to-RBC ratio of 2:1 is more effective in correcting trauma-induced coagulopathy than a ratio of 1:1 in a rat multiple trauma transfusion model

    DOI: 10.1186/s40635-019-0242-5
    Sample: Blood
  4. Experimental and Therapeutic Medicine (2017) IF: 1.261
    Sivelestat sodium hydrate improves post‑traumatic knee osteoarthritis through nuclear factor‑κB in a rat model

    DOI: 10.3892/etm.2017.4684
    Sample: Serum
  5. Experimental and Therapeutic Medicine (2020) IF: 1.785
    Human umbilical cord‑derived mesenchymal stem cells and human cord blood mononuclear cells protect against cisplatin‑induced acute kidney injury in rat models

    DOI: 10.3892/etm.2020.9274
    Sample: Tissue Homogenate
  6. MEDICAL SCIENCE MONITOR (2020) IF: 1.918
    Carbon Monoxide Inhibits the Expression of Proteins Associated with Intestinal Mucosal Pyroptosis in a Rat Model of Sepsis Induced by Cecal Ligation and Puncture

    DOI: 10.12659/MSM.920668
    Sample: Serum,Intestinal Tissue
  7. Pregnancy Hypertension-An International Journal of Womens Cardiovascular Health (2020) IF: 2.095
    Glycyrrhizin potentially suppresses the inflammatory response in preeclampsia rat model

    DOI: 10.1016/j.preghy.2020.10.007
    Sample: Serum
  8. PLoS One (2022) IF: 3.24
    PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells

    DOI: 10.1371/journal.pone.0265191

    PMID: 35294955

    		 Sample: Vsmcs
  9. Journal of Trauma and Acute Care Surgery (2020) IF: 3.381
    Impact of uric acid on liver injury and intestinal permeability following resuscitated hemorrhagic shock in rats

    DOI: 10.1097/TA.0000000000002868

    PMID: 33231951

    		 Sample: Plasma
  10. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY (2019) IF: 3.553
    HMGB1 is mechanistically essential in the development of experimental pulmonary hypertension

    DOI: 10.1152/ajpcell.00148.2018

    PMID: 30517029

    		 Sample: Plasma,Cell Culture Supernatant
  11. TOXICOLOGY (2015) IF: 3.621
    Ethanol directly induced HMGB1 release through NOX2/NLRP1 inflammasome in neuronal cells

    DOI: 10.1016/j.tox.2015.06.006
    Sample: Cell Culture Supernatant
  12. BIOMEDICINE & PHARMACOTHERAPY (2019) IF: 3.743
    HMGB1/RAGE pro-inflammatory axis promotes vascular endothelial cell apoptosis in limb ischemia/reperfusion injury

    DOI: 10.1016/j.biopha.2019.109005
    Sample: Serum,Human Umbilical Vein Endothelial Cells (Huve
  13. Drug Design Development and Therapy (2021) IF: 4.162
    Prunella vulgaris L. Attenuates Experimental Autoimmune Thyroiditis by Inhibiting HMGB1/TLR9 Signaling

    DOI: 10.2147/DDDT.S325814

    PMID: 34764638

    		 Sample: Blood
  14. Scientific Reports (2021) IF: 4.38
    Protective effect of glycyrrhizin, a direct HMGB1 inhibitor, on post-contrast acute kidney injury

    DOI: 10.1038/s41598-021-94928-5

    PMID: 34341389

    		 Sample: Serum
  15. ADDICTION BIOLOGY (2016) IF: 4.547
    Oleoylethanolamide prevents neuroimmune HMGB1/TLR4/NF‐kB danger signaling in rat frontal cortex and depressive‐like behavior induced by ethanol binge administration

    DOI: 10.1111/adb.12365
    Sample: Plasma
  16. Pharmaceuticals (2022) IF: 5.215
    Repositioning Linagliptin for the Mitigation of Cadmium-Induced Testicular Dysfunction in Rats: Targeting HMGB1/TLR4/NLRP3 Axis and Autophagy

    DOI: 10.3390/ph15070852

    PMID: 35890148

    		 Sample: Testicular Tissue
  17. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (2021) IF: 5.31
    Normothermic machine perfusion attenuates hepatic ischaemia-reperfusion injury by inhibiting CIRP-mediated oxidative stress and mitochondrial fission

    DOI: 10.1111/jcmm.17062

    PMID: 34786826

    		 Sample: Serum
  18. LIFE SCIENCES (2022) IF: 6.78
    Irbesartan reprofiling for the amelioration of ethanol-induced gastric mucosal injury in rats: Role of inflammation, apoptosis, and autophagy

    DOI: 10.1016/j.lfs.2022.120939

    PMID: 36115582

    		 Sample: Gastric Tissue
  19. Cells (2023) IF: 7.666
    Azilsartan Modulates HMGB1/NF-κB/p38/ERK1/2/JNK and Apoptosis Pathways during Renal Ischemia Reperfusion Injury

    DOI: 10.3390/cells12010185
    Sample: Kidney Tissue
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Reviews/Q&A

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  • Reviews
  • Q&A
Q

M***aSubmitted [ Jul 06 2018 ]

Asked: Hi! I saw in the protocol of this kit that it only works with EDTA and heparin as anticoagulant. Sometimes, although protocols don\'t say it, this kits also works with citrate as anticoagulant. I would like to know whether or not this is the case. Furthermore, I would like to know how much time would it take to the shipping to arrive to Madrid (Spain). Thank you very much, Marta

Reply

A

adminSubmitted [ Jul 06 2018 ]

Answered: Dear Marta, Thanks for your question. This kit could test with citrate plasma, but according to our experiments, the accuracy of the results decreased slightly. So we recommend to use EDTA and heparin as anticoagulant. It usually takes 3~5 days to ship the kits to arrive in Madrid (Spain). Please feel free to contact us if any question. Thanks!

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