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

  • Cat.No.:E-EL-R0505

  • Reactivity: Rat

To Purchase E-EL-R0505

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609
Qty:

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℃, 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

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

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

  1. Addiction Biology (2017) IF: 4.603
    Oleoylethanolamide Prevents Neuroimmune HMGB1/TLR4/NF-κB Danger Signaling In Rat Frontal Cortex And Depressive-Like Behavior Induced By Ethanol Binge Administration.

    DOI: 10.1111/adb.12365

    PMID: 26857094

    Sample: Plasma
  2. Addiction Biology (2016) IF: 4.603
    Oleoylethanolamide Prevents Neuroimmune HMGB1/TLR4/NF-κB Danger Signaling In Rat Frontal Cortex And Depressive-Like Behavior Induced By Ethanol Binge Administration.

    DOI: 10.1111/adb.12365

    PMID: 26857094

    Sample: Plasma
  3. American Journal of Physiology-Cell Physiology (2018) IF: 3.454
    HMGB1 is mechanistically essential in the development of experimental pulmonary hypertension

    DOI: 10.1152/ajpcell.00148.2018

    PMID: 30517029

    Sample: Plasma,Cell culture supernatant
  4. Toxicology (2015) IF: 3.582
    Ethanol Directly Induced HMGB1 Release Through NOX2/NLRP1 Inflammasome In Neuronal Cells.

    DOI: 10.1016/j.tox.2015.06.006

    PMID: 26079697

    Sample: Cell culture supernatant
  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. Experimental and Therapeutic Medicine (2017) IF: 1.261
    Sivelestat Sodium Hydrate Improves PostTraumatic Knee Osteoarthritis Through Nuclear FactorκB In A Rat Model.

    DOI: 10.3892/etm.2017.4684

    PMID: 28810618

    Sample: Serum
  7. United States Patent Application Publication (2018)
    Liposomal rehydration salt formulation for treatment of alcohol related disorders
    Sample: Plasma
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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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