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

Cat.No.:E-EL-R0505
Reactivity: Rat

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48T
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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.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 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), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 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

Synonyms	HMGB1, HMG1, HMG3, SBP-1, Amphoterin
Research Area	Cancer, Epigenetics and Nuclear Signaling

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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

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Reviews/Q&A

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Reviews
Q&A

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

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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