Overview

  • Product name

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

    Colorimetric
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 5 pg/ml
  • Range

    2.74 pg/ml - 2000 pg/ml
  • Recovery

    100 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 102.34 90% - 111%
    Tissue Extracts 98.26 89% - 109%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

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


    This assay employs an antibody specific for mouse 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-mouse 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.


    Get higher sensitivity in only 90 minutes with Mouse IL-1 beta ELISA Kit (ab197742) from our SimpleStep ELISA® range.


     

  • 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
    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-mouse IL-1 beta 2 vials
    IL-1 beta Microplate (12 x 8 wells) 1 unit
    Recombinant Mouse IL-1 beta Standard (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

Associated products

Applications

Our Abpromise guarantee covers the use of ab100705 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

  • Dilution curves of mouse IL-1 beta (ab100705) and NIBSC standard (93/668). One ng of standard mouse IL-1 beta corresponds to 34.6 (+/-2.1) AU NIBSC 93/668. Background signal subtracted (duplicates; +/- SD).

  • IL-1 beta measured in mouse tissue lysates (0.02-0.3 mg x mL-1 protein tested, data expressed per

    mg of extracted protein; duplicates +/- SD).

  • IL-1 beta measured in lysates from control cells, or cells stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297), with the addition of 1 ug x mL-1 of LPS (Sigma) (P/L) for the last 6 hours (duplicates +/- SD).

  • Representative standard curve using ab100705

Protocols

References

This product has been referenced in:

  • 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) »
  • Lai XL  et al. Apc gene suppresses intracranial aneurysm formation and rupture through inhibiting the NF-?B signaling pathway mediated inflammatory response. Biosci Rep 39:N/A (2019). Read more (PubMed: 30808715) »
See all 10 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

GOOD

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Sample: Mouse Cell (Homogenised lung tissue)
Specification: Homogenised lung tissue
Type of Kit Used: ELISA
Positive control: Virally infected lung tissue.
Did you use the Abcam Kit protocol?: Yes
Additional data
Additional Notes: Overall, a good kit.

Abcam user community

Verified customer

Submitted Jan 27 2016

Answer



It is possible to use the non-lysate kit ab100704 (IL-1 beta (Interleukin-1 beta) Mouse ELISA Kit) and use your own lysis buffers (non-denaturing, with a mild detergent like RIPA/NP40/Triton X). The cell lysate kit (ab100705) contains this buffer and a different assay diluent, but the different assay diluent is not required to run the assay correctly with tissue/cell lysate samples.

I can recommend to use a detergent based on our sample preparation tips here:

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. 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

ıl1beta mouse elisa kit ab100705

Average Good 4/5 (Ease of Use)
Abreviews
This kit ( ab100705 ) is not suitable for the detection of il1 beta levels from the serum of mice. We collected blood from ApoE-/- mice that are on high fat diet for 3 months in addition to ApoE-/- mice fed with chow diet. In none of these serum samples we obtained a promising result. On the other hand, the same kit detected il1beta from liver lysates as kit suggested.

Abcam user community

Verified customer

Submitted Jun 29 2015

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 your reply.


Because we carry over 100,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.


Regrettably, as we haven't tested or even compared the reactivity to the pro- or mature form of the IL1 beta, we cannot guarantee it will work with both forms.

However, we are still waiting for a response from our collaborator of ab108866. So there is still a change that the reactivity has been measured.


I hope this information is still helpful. We will let you know as soon as we got a reply.

Read More

Answer

Thank you for your patience.

I am still waiting on more information on ab108866.

But I am happy to provide the following information about ab100704 and ab100705:

The antibody pair used in the Mouse IL-1 beta Sandwich ELISA kits was raised against an immunogen which was the mature/cleaved form of the IL-1 beta protein consisting of amino acids 118-269. Therefore we know that the kit can detect the active/mature form.

Whether the kit can also detect the propeptide however has not been determined so accordingly we don’t know if this kit can differentiate between the propeptide and the mature form. The only way to determine this would be to do either epitope mapping (which we haven’t done as it is time-consuming and costly) or to simply spike in only the propeptide and see if the kit detects it.

Because the propeptide does contain the amino acid sequence of the mature peptide though, if it’s a linear epitope then likely the kit would also detect the propeptide. If the epitope is confirmational or some post-translational modification blocks the binding site, then the kit would likely not detect the propeptide). This is all speculation of course because we don’t know the epitope.

I hope this information is helpful. Please do not hesitate to contact em again with any further questions.

Read More

Abreview for IL1 beta Mouse ELISA Kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews

Sample: Mouse Purified protein (Placenta tissue protein)
Specification: Placenta tissue protein
Amount of Sample: 0.4 µg
Type of Kit Used: ELISA
Did you use the Abcam Kit protocol?: Yes

Additional data
Additional Notes: Bar-1, control
Bar-2, LPS
Bar-3, Surfactant Protein-A
Bar-4, LPS + SP-A

Dr. Varkha Agrawal

Verified customer

Submitted Sep 26 2012

Question
Answer

Thank you for your call yesterday and for your patience while I've been in touch with the lab regarding your enquiry.

We are still looking into the question of what kind of tissue ab100705 was tested with, and I will be back in touch with this information. However I did obtain some suggestions for using ab100747 and ab1007390 with tissue lysates, and I wanted to forward this on to you.

Our kits are very accommodating to many different samples types so lysates will likelywork, though we can't guarantee it.When testing a kit with tissue lysates, we recommend diluting the samples at least 5-fold with the diluent or cell culture media to minimize any effects of the detergents in the lysis buffer.The samples may need to be diluted further but this would need to be determined empirically.

In brief, a lysis buffer must meet the following specifications:


A) has relatively low salt content (700 mM or less)
B) does not contain sodium azide
C) does not contain >0.1% SDS
D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)

This would include any buffers used for immunoprecipitations, including RIPA buffer.
I hope this information will be useful, but please let me know if you have any further questions or if there is anything else that we can do for you. I'll be back in touch once I have more information about ab100705.

Read More

Answer

Thank you for your email.
I can confirm that this kit will recognize the mature form (aa 118-269) of IL1beat.
Since the pro IL1beta also contains this amino acid sequence, I would expect the kit to also detect the pro-form. This has not been experimentally tested by us.
The IL1 beta Mouse ELISA Kit does not cross-react with IL1alpha. This has been experimentally tested by the lab.
I have to confirm that ab100705 has not yet been referenced specifically in any publications we know of.
I copied our suggestions for the preparation of tissue lysates below.
I hope this information is helpful. Please do not hesitate to contact me again with any further questions.


How do I make cell or tissue lysates for use on the cytokine arrays?

The cell or tissue lysates for use with ab100705 ELISA kit can be prepared using most conventional methods, e.g. homogenization of cell or tissue in Lysis Buffer. We supply a compatible lysis buffer in most of our kits. Other general low-salt lysis buffers can be used with the following caveats:
1) Avoid using 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 reducing agents, such as dithiothreitol or mercaptoethanols
In general, we strongly recommend that you add some type of protein inhibitor “cocktail” to the lysis buffer prior to homogenization. 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 protein 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 (activated) forms of the protein.
Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freeze-thaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no one “best method” for all sample types, but some are better than others for some 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, spin down the lysates to remove cell/tissue debris (5 min @ 10000 x g or 10 min @ 5000 x g) and save the supernatant. Lysates should be frozen as soon as possible, and stored at -20°C (or -80°C, if possible). Spin them again before incubating with the antibody array. Determine the protein concentration of your lysates (using a Bradford-Lowry-type assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay.
Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 uL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target for total protein concentration of the homogenate should be at least 1000 ug/mL, but 2000 ug/mL would be better.

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