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    rat-il-1-beta-elisa-kit-ab100768.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Rat IL-1 beta ELISA Kit (ab100768)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (5)References (41)

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Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)
  • Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)
  • Sandwich ELISA - IL-1 beta (Interleukin-1 beta) Rat ELISA Kit (ab100768)
  • Typical Standard Curve

Key features and details

  • Sensitivity: 80 pg/ml
  • Range: 68.59 pg/ml - 50000 pg/ml
  • Sample type: Cell culture extracts, Tissue Extracts
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Rat

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Overview

  • Product name

    Rat IL-1 beta ELISA Kit
    See all IL-1 beta kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall < 10%
    Inter-assay
    Sample n Mean SD CV%
    Overall < 12%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 80 pg/ml
  • Range

    68.59 pg/ml - 50000 pg/ml
  • Recovery

    116 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 96.22 88% - 105%
    Tissue Extracts 136.5 126% - 145%
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Rat
  • Product overview

    Abcam’s IL-1 beta Rat ELISA (Enzyme-Linked Immunosorbent Assay) Kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of rat IL-1 beta in cell lysates and tissue lysates.


    This assay employs an antibody specific for rat IL-1 beta coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-1 beta present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-rat IL-1 beta antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and colour develops in proportion to the amount of IL-1 beta bound. The Stop Solution changes the colour from blue to yellow, and the intensity of the colour is measured at 450 nm.

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysis Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    5X Sample Diluent Buffer 1 x 10ml
    Biotinylated anti-rat IL-1 beta 2 vials
    IL-1 beta Microplate (12 x 8 wells) 1 unit
    Recombinant rat IL-1 beta Standard (lyophilized)  2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
    • Metabolism
    • Types of disease
    • Obesity
  • Function

    Potent proinflammatory cytokine. Initially discovered as the major endogenous pyrogen, induces prostaglandin synthesis, neutrophil influx and activation, T-cell activation and cytokine production, B-cell activation and antibody production, and fibroblast proliferation and collagen production. Promotes Th17 differentiation of T-cells.
  • Tissue specificity

    Expressed in activated monocytes/macrophages (at protein level).
  • Sequence similarities

    Belongs to the IL-1 family.
  • Post-translational
    modifications

    Activation of the IL1B precursor involves a CASP1-catalyzed proteolytic cleavage. Processing and secretion are temporarily associated.
  • Cellular localization

    Cytoplasm, cytosol. Lysosome. Secreted, exosome. Cytoplasmic vesicle, autophagosome. Secreted. The precursor is cytosolic. In response to inflammasome-activating signals, such as ATP for NLRP3 inflammasome or bacterial flagellin for NLRC4 inflammasome, cleaved and secreted. IL1B lacks any known signal sequence and the pathway(s) of its secretion is(are) not yet fully understood (PubMed:24201029). On the basis of experimental results, several unconventional secretion mechanisms have been proposed. 1. Secretion via secretory lysosomes: a fraction of CASP1 and IL1B precursor may be incorporated, by a yet undefined mechanism, into secretory lysosomes that undergo Ca(2+)-dependent exocytosis with release of mature IL1B (PubMed:15192144). 2. Secretory autophagy: IL1B-containing autophagosomes may fuse with endosomes or multivesicular bodies (MVBs) and then merge with the plasma membrane releasing soluble IL1B or IL1B-containing exosomes (PubMed:24201029). However, autophagy impacts IL1B production at several levels and its role in secretion is still controversial. 3. Secretion via exosomes: ATP-activation of P2RX7 leads to the formation of MVBs containing exosomes with entrapped IL1B, CASP1 and other inflammasome components. These MVBs undergo exocytosis with the release of exosomes. The release of soluble IL1B occurs after the lysis of exosome membranes (By similarity). 4. Secretion by microvesicle shedding: activation of the ATP receptor P2RX7 may induce an immediate shedding of membrane-derived microvesicles containing IL1B and possibly inflammasome components. The cytokine is then released in the extracellular compartment after microvesicle lysis (PubMed:11728343). 5. Release by translocation through permeabilized plasma membrane. This may occur in cells undergoing pyroptosis due to sustained activation of the inflammasome (By similarity). These mechanisms may not be not mutually exclusive.
  • Target information above from: UniProt accession P01584 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Catabolin
    • H1
    • IFN beta inducing factor
    • IL 1
    • IL 1 beta
    • IL-1 beta
    • IL1
    • IL1 BETA
    • IL1B
    • IL1B_HUMAN
    • IL1F2
    • Interleukin 1 beta
    • Interleukin 1 beta precursor
    • interleukin 1, beta
    • Interleukin-1 beta
    • OAF
    • Osteoclast activating factor
    • OTTHUMP00000162031
    • Preinterleukin 1 beta
    • Preinterleukin beta
    • Pro interleukin 1 beta
    see all
  • Database links

    • Entrez Gene: 24494 Rat
    • SwissProt: Q63264 Rat
    • Unigene: 9869 Rat

    Associated products

      Images

      • Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)
        Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)

        Standard curve of Rt IL-1 beta, with background signal subtracted (duplicates; +/- SD).

      • Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)
        Sandwich ELISA - IL-1 beta Rat ELISA Kit (ab100768)
        Rat IL-1b measured in biological fluids or in spleen lysate; background signal subtracted (duplicates +/- SD).
      • Sandwich ELISA - IL-1 beta (Interleukin-1 beta) Rat ELISA Kit (ab100768)
        Sandwich ELISA - IL-1 beta (Interleukin-1 beta) Rat ELISA Kit (ab100768)
        Rat IL-1b detected in cell lysates from NR8383.1 control cells (C) or cells stimulated for 6 hours with 5 ug x mL-1 of LPS (Sigma). Results shown for IL-1? levels in 10e6 cells, after background signal was subtracted (duplicates +/- SD).
      • Typical Standard Curve
        Typical Standard Curve

        Representative standard curve using ab100768

      Protocols

      • Protocol Booklet

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
      • SDS
    • References (41)

      Publishing research using ab100768? Please let us know so that we can cite the reference in this datasheet.

      ab100768 has been referenced in 41 publications.

      • Zhao Q  et al. Chitoheptaose Promotes Heart Rehabilitation in a Rat Myocarditis Model by Improving Antioxidant, Anti-Inflammatory, and Antiapoptotic Properties. Oxid Med Cell Longev 2020:2394704 (2020). PubMed: 32351668
      • Wang Y  et al. miR-27a suppresses TLR4-induced renal ischemia-reperfusion injury. Mol Med Rep 20:967-976 (2019). PubMed: 31173204
      • Sun L  et al. The involvement of spinal annexin A10/NF-?B/MMP-9 pathway in the development of neuropathic pain in rats. BMC Neurosci 20:28 (2019). PubMed: 31208343
      • Sun J  et al. Cytoprotective effects of galacto-oligosaccharides on colon epithelial cells via up-regulating miR-19b. Life Sci 231:116589 (2019). PubMed: 31226416
      • Hellenbrand DJ  et al. Sustained interleukin-10 delivery reduces inflammation and improves motor function after spinal cord injury. J Neuroinflammation 16:93 (2019). PubMed: 31039819
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-6 of 6 Abreviews or Q&A

      Question

      We would like to assess both IL-1 Beta and TNF alpha concentration in homogenate of colonic tissue, but in the data sheet is not specified the procedure to obtain the tissue homogenate. Do you have available a standardized extraction methodology? or could you suggest any references describing such a procedures?

      Read More

      Abcam community

      Verified customer

      Asked on Mar 12 2014

      Answer



      I can suggest the following general tissue lysate preparation procedure to prepare samples for both the kits.




      Tissue lyste samples can be prepared using most conventional methods, for example with RIPA or other formulations.

      Please note the following guidelines on lysis buffer composition:
      1) Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic
      detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as
      CHAPS, or mild ionic detergents such as sodium deoxycholate will work.
      2) Use no more than 2% v/v total detergent
      3) Avoid the use of sodium azide
      4) Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols

      We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to
      homogenization. Most general biochemical supply companies including stock a wide variety of these products. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used but are not necessary unless the antibodies used in the kit specifically
      recognize phosphorylated forms of the protein.
      Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freezethaw,
      sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these.
      There is no best method for all sample types; your choice of method should be made following a brief
      search of the literature to see how samples similar to yours have been prepared in previous
      investigations.
      After homogenization, centrifuge the lysates to remove cell/tissue debris (5 min @ 10,000 x g or 10
      min @ 5,000 x g) and save the supernatant. Unless testing fresh, lysates should be frozen as soon as
      possible and stored at -20°C (or -80°C, if possible). Centrifuge them again before incubating with any
      immunoassay. Next, determine the protein concentration of your lysates using a total protein assay
      not inhibited by detergents (such as the Bicinchoninic acid (BCA) assay) and normalize the volume of
      each sample used to deliver the same amount of total protein for each assay.

      Note: The Bradford assay is not recommended as it can be inhibited by the presence of detergents.
      Since different cells and tissues may contain different amounts of protein, as starting point, we
      suggest using 500 μL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this
      based upon your results. Your target total protein concentration of the homogenate should be at least
      1,000 μg/mL, but 2,000 μg/mL or more would be better.

      Read More

      Sam Washer

      Abcam Scientific Support

      Answered on Mar 12 2014

      Testing IL-1beta levels in rat spinal cord.

      Below average Average 3/5 (Ease of Use)
      Abreviews
      Abreviews
      abreview image
      We tested IL-1β levels in homogenized rat spinal cord. The results had to high of background and the CV values were extremely high. The tech help from ABCAM was very helpful. We tried several of their recommendations including using a blocker, different dilutions, centrifuging again before adding samples, and triplicate wells, but nothing helped. It’s possible this will work for other sample types, but I would not recommend it for spinal cord. Note the high CV values in the image.

      Abcam user community

      Verified customer

      Submitted Mar 15 2020

      Question

      I´m interested to measure IL-1 beta (Rat IL-1 beta ELISA Kit (Interleukin-1 beta) (ab100768)) and TNF (Rat TNF alpha ELISA Kit (ab100785) levels in rat hippocampus tissue. I've read the manual and it says that the kit can measure these cytokines levels (tissue homogenates),however it is not specified in what kind of buffer it must be homegenize the tissues. My question is: Can I use these Cytokines kits using homgenates in Lysis buffer ( 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0)?

      Read More

      Abcam community

      Verified customer

      Asked on Jan 16 2017

      Answer

      Yes, that lysis buffer will be good, the 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0.


      Here are some general guidelines.


      Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work. 


      Use no more than 2% v/v total detergent  


      Avoid the use of sodium azide 


      Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols 


      We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization.

      Read More

      Abcam Scientific Support

      Answered on Jan 16 2017

      Question

      herzlichen Dank für Ihre schnelle Antwort.

      Die Tiere werden mit KH-Medium (Krebs-Henseleit-Medium) perfundiert.Das besteht aus vier verschiedenen Stocklösungen, die aus NaCl, KCl, KH2PO4, MgSO4 x 7H2O, NaHCO3 und CaCl2 x 2H2O hergestellt werden, also ganz normale Salzlösungen.

      Vielen Dank und mit freundlichen Grüßen,

      Read More

      Abcam community

      Verified customer

      Asked on Oct 22 2012

      Answer



      Da die Tiere ja nur "durchgespült", aber nicht fixiert wurden, sollte dieses Kit mit den Leberlysaten funktionieren.

      Read More

      Abcam Scientific Support

      Answered on Oct 22 2012

      Question

      in unserer Arbeitsgruppe beschäftigen wir uns mit chronischen Lebererkrankungen und im Rahmen unseres aktuellen Projekts möchten wir gerne IL-1 beta messen. Die Proben, die wir messen möchten, sind Perfusate aus der Rattenleber.
      Meine Frage wäre jetzt, ob es mit diesem ELISA Kit (ab100768) möglich ist, unsere Perfusate zu messen?

      Herzlichen Dank und mit freundlichen Grüßen,

      Read More

      Abcam community

      Verified customer

      Asked on Oct 19 2012

      Answer

      Vielen Dank für Ihre Anfrage.

      Womit perfusieren Sie den die Tiere, PFA oder ein anderes Fixierungsmittel oder einfach nur PBS?

      Ich freue mich auf Ihre Antwort.

      Read More

      Abcam Scientific Support

      Answered on Oct 19 2012

      Question

      Dear Technical team, Our user ordered this product. She wants to test Rat Plasma, unfortuntely could not confirm in your data sheet. Could you please check this for our user? I look forward to hearing from you. Best regards,

      Read More

      Abcam community

      Verified customer

      Asked on Sep 01 2011

      Answer

      Thank you for your enquiry. I can confirm that ab100767 IL1 beta Rat ELISA Kit would be the suitable kit to use with plasma samples. ab100768 IL1 beta Rat ELISA Kit is not designed for use with plasma samples. I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

      Read More

      Abcam Scientific Support

      Answered on Sep 01 2011

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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