Overview

  • Product name

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

    Colorimetric
  • Sample type

    Cell culture supernatant, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 0.3 pg/ml
  • Range

    0.48 pg/ml - 100 pg/ml
  • Recovery

    99 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 100.43 89% - 110%
    Plasma 99.66 90% - 107%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human IL-1 beta ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IL-1 beta in plasma and cell culture supernatants. (Human IL-1 beta concentration is quite low in normal plasma, it may not be detected in this assay).


    This assay employs an antibody specific for human 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-Human 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 color develops in proportion to the amount of IL-1 beta bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    We have not been able to detect the endogenous human IL-1 beta in normal serum with ab100562, only in serum spiked with human IL-1 beta.

  • Notes

    Optimization may be required with urine samples.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    300X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human IL-1 beta  2 vials
    IL-1 beta Microplate (12 x 8 wells) 1 unit
    Recombinant Human IL-1 beta Standards (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • 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.
  • 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

Applications

Our Abpromise guarantee covers the use of ab100562 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Hu IL-1 beta measured in biological fluids showing quantity (pg) per mL of tested sample (U937: stimulated 24h with 10ng/mL PMA and 6h with LPS 1microgram/mL -- THP-1: stimulated 24h with 10ng/mL PMA)

  • Representative standard curve using ab100562

  • Representative standard curve using ab100562

Protocols

References

This product has been referenced in:

  • Ran Z  et al. Curcumin inhibits high glucose-induced inflammatory injury in human retinal pigment epithelial cells through the ROS-PI3K/AKT/mTOR signaling pathway. Mol Med Rep 19:1024-1031 (2019). Read more (PubMed: 30569107) »
  • Turner CT  et al. Granzyme K Expressed by Classically Activated Macrophages Contributes to Inflammation and Impaired Remodeling. J Invest Dermatol 139:930-939 (2019). Read more (PubMed: 30395844) »
See all 13 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Answer

Gracias por contactarnos.


He querido comprobar la posibilidad de presentar reactividad cruzada entre ratón y humano en los kits contra las citoquinas que mencionas.

Tenemos varios kits contra dichas citoquinas específicos para ratón, y específicos para humano. Después de estudiar la homología entre secuencias, me temo que no podemos predecir que exista reactividad cruzada entre especies. Experimentalmente tampoco tenemos datos para confirmarlo.

Siento no poder ser de mas ayuda. No dudes en contactarnos para cualquier otra consulta.

Read More

Answer

Thank you for contacting us.

You will need 100ul of antibody for each well in 96 well plate. Say you are going to use all 96 wells so you will need final volume of antibody 9600ul (9.6 ml).

There is always a loss of few micro litres while pipetting so we recommend preparing more than what is required. e.g. add 14 ul of antibody from step 6 to 1106 ul of assay diluent B. This will give you total of 1120 ul i.e. 80 times diluted antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Which buffer you have to use depends on what kind of sample you are using. What kind of sample are you using?

Assay Diluent B should be used with the antibodies as Assay Diluent A contains 0.09% sodium azide as
preservative.This may interfere with the HRP conjugate. However, I will check with the lab if this is correct.


I am looking forward to your reply with the requested details.

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Question
Answer

Yes, It is all free however they need to check if the items were paid in original order.

It is just a normal check so there is no need to worry, you will be getting a order confirmation soon.

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Answer

Our accountsdepartment is holding the order at the moment. The order will be shipped once they authorize it, after checking thepayment etc. Please email mailto:creditcontrol@abcam.com for any order update.

Thanks!

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Answer

Thank you for contacting us.

To prevent the shortageissue in the future, please advise your customer along with other customers to check all reagents before beginning the experiment and that if there is a volume shortage or any other issue to not try and use this reagents without first contacting us to prevent what is most of the time an easily correctable and avoidable problem.

We have place order for 2 kits ab100562that will be shipped to you soon. The order number is 1130294.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your cooperation.

I have sent a request to lab for new kits and hoping to get an answer later this evening. I will confirm you the dispatch date.

I hope this info will be helpful.

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Answer

Thank you for your email.

Regarding ab100562 have you used both kits? If not, then I will send you one replacement kit and one extra vial of HRP-standard. Could you let me know if you still have to use one kit of ab100562?

There are many factor that affect the standard curve. Could you please check the following when doing experiment;

- the reagents should be at room temperature when used
- avoid any pipetting error
- keep TMB substrate away from light
- check the ELISA reader wavelengths
- check standard is properly reconstituted and mixed
- try incubating the primary antibody for 24 hours.

I am sure these tips can help you get good results. Please do not hesitate to contact me if results does not improve.

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Answer

Thank you for your email. I apologize for any inconvenience caused.

The volume 200ul is actually more than what is need for the assay; this extra volume is added to avoid any loss due to evaporation, insufficient centrifuging, etc. Actually only 32ul X 300ul = 9.6 ml is needed for full 96 well assay plate.

I have personally checked the in-stock Item G of same lot which you have received. The volume was ˜50 ul, I accept it is not 200ul so I have informed our lab to tighten the caps of all vials. 10ul as you mentioned is quite less so I would like to ask, if you spin the vial in centrifuge before taking the liquid out? We always recommend our customers spinning the vials, in order to recover maximum amount possible.

Could you tell us more about the problem? Was your experiment failed due to insufficient HRP-strepatividin or due to other factors. Did you get standard curve? The recovered 10ul solution would have been enough for 25-30 wells; did you not get good results?

The kit ab100529 can detect GM-CSF in range between 2-500 picogramm/ml.

The standard should be made by the end user. Please check the protocol booklet for details about standard curve and detection range.

I hope this information will be helpful. I will be looking forward to hearing from you soon.

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1-10 of 15 Abreviews or Q&A

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