Overview

  • Product name
  • Description
    Rabbit polyclonal to IL-1 beta
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ELISA, IHC-P, RIA, IHC-Fr, Functional Studies, Flow Cyt, Neutralising, ICC/IF, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Non human primates
  • Immunogen

    Recombinant 153 aa human IL-1beta produced in E.coli. MW of recombinant IL-1beta 17kDa, with the N-terminal amino acid at position alanine 117. This is the cleavage site generated by the IL-1beta converting enzyme (ICE, capase-1).

  • General notes
    Immunohistochemistry 1/100 to 1/200 (paraffin or cryofixation can be used; 1/100 for staining intracellular IL-1beta). ELISA 1/200 to 1/1000 (this antibody is best used as the second antibody with a monoclonal as a capture antibody). Radio immunoassay 1/8000 Neutralization of IL-1beta activity in bioassays 1/100 ( >4 hours incubation, normal rabbit IgG as negative control) FACS analysis - caution, F(c) domain of rabbit IgG may interact with cells non-specifically.

Properties

Applications

Our Abpromise guarantee covers the use of ab2105 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
WB Use at an assay dependent concentration.

1/1000 - 1/2000 (antigen - supernatants or lysates of 2 x 106 endotoxin-stimulated human PBMC. Denatured 31kDa precursor IL-1beta will be recognized, but often migrates as a 35 kDa band).

IP Use at an assay dependent concentration. 1/400 - 1/800 (pre-clearing with a non-specific rabbit IgG is helpful to reduce background).
ELISA 1/500 - 1/2000.
IHC-P 1/100 - 1/200.
RIA 1/8000.
IHC-Fr 1/100 - 1/200.
Functional Studies Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

F(c) domain of rabbit IgG may interact with cells non-specifically!

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Neutralising 1/100. Neutralization of IL-1beta activity in bioassays 1/100 ( >4 hours incubation, normal rabbit IgG as negative control). ab2105 does not neutralize the biological activity of murine, rat or rabbit IL1 beta.
ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 28726778

Target

  • 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
  • Database links
  • Alternative names
    • Catabolin antibody
    • H1 antibody
    • IL 1 antibody
    • IL 1 beta antibody
    • IL-1 beta antibody
    • IL1 BETA antibody
    • IL1B antibody
    • IL1B_HUMAN antibody
    • IL1F2 antibody
    • Interleukin 1 beta antibody
    • Interleukin-1 beta antibody
    • OAF antibody
    • OTTHUMP00000162031 antibody
    • Preinterleukin 1 beta antibody
    • Pro interleukin 1 beta antibody
    see all

Images

  • All lanes : Anti-IL-1 beta antibody (ab2105) at 1/1000 dilution

    Lane 1 : Human peripheral blood mononuclear cell lysate - PBMC's unstimulated
    Lane 2 : Human peripheral blood mononuclear cell lysate - monocytes unstimulated
    Lane 3 : Human peripheral blood mononuclear cell lysate - monocytes stimulated with LPS
    Lane 4 : Human peripheral blood mononuclear cell lysate - positive control (rh IL-1 beta protein)

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution

    Performed under reducing conditions.

    Observed band size: 35 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 minutes

    See Abreview

  • ab2105 at 1/20 staining human liver tissue sections by IHC-P. The cells were paraformaldehyde fixed and a heat mediated antigen retrieval step was performed. The tissue was incubated with the tissue overnight at room temperature. A biotinylated donkey anti-rabbit IgG was used as the secondary antibody and this was detected using streptavidin HRP.

    See Abreview

  • ab2105 staining IL-1 beta in human keratinocytes by Immunocytochemistry/ Immunofluorescence. Cells were fixed in methanol, permeabilized in 0,1% Saponin / PBS and then blocked using 4% BSA for 30 minutes at 25°C. Samples were then incubated with ab2105 at a 1/100 dilution for 2 hours at 4°C. An Alexa-fluor 488 conjugated goat anti-rabbit polyclonal was used as the secondary antibody at a 1/100 dilution.

    See Abreview

  • ab2105 staining IL-1 beta in Human monocytes by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with 0.1% saponin in PBS. The sample was incubated with the primary antibody (1/500 in 0.1% saponin + 1% FCS in PBS) for 45 minutes at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG (1/100) was used as the secondary antibody.

    1 - unstimulated monocytes. 2 - monocytes stimulated with LPS.

    See Abreview

  • Lanes 2-5 : Anti-IL-1 beta antibody (ab2105) at 1/1000 dilution

    Lane 1 : Ladder (top to bottom: p35, p25, p20, p17, p11)
    Lane 2 : Human THP-1 leukemic monocyte cell lysate, no LPS at 20 µg
    Lane 3 : Human THP-1 leukemic monocyte supernatant, no LPS at 20 µg
    Lane 4 : Human THP-1 leukemic monocyte cell lysate, LPS stimulation at 20 µg
    Lane 5 : Human THP-1 leukemic monocyte supernatant, LPS stimulation at 20 µg

    Secondary
    Lanes 2-5 : IRDye® 800CW-conjugated Donkey anti-rabbit IgG monoclonal at 1/5000 dilution

    Performed under reducing conditions.

    Observed band size: 17,30 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Both mature and pro-IL-1beta bands, p17 and p31 were detected by ab2105 in supernatant from THP-1 cells stimulated with LPS. Only pro-IL-1beta band (p31) was detected in supernatant from THP-1 cells not stimulated with LPS.

    See Abreview

References

This product has been referenced in:
  • Wang S  et al. Oxidative stress, autophagy and pyroptosis in the neovascularization of oxygen-induced retinopathy in mice. Mol Med Rep 19:927-934 (2019). Read more (PubMed: 30569132) »
  • Wang X  et al. Effect of Gastrodin on Early Brain Injury and Neurological Outcome After Subarachnoid Hemorrhage in Rats. Neurosci Bull N/A:N/A (2019). Read more (PubMed: 30673960) »
See all 57 Publications for this product

Customer reviews and Q&As

21-30 of 38 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Shoulder tissue)
Antigen retrieval step
None
Permeabilization
No
Specification
Shoulder tissue
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Formaldehyde

Ms. Mindy Cote

Verified customer

Submitted Dec 15 2011

Answer

I have talked to some of my colleagues and the only things we can think of are, things that I am sure you have already thought about but, first the most basic: Be sure the CD11 antibody is non-rabbit. If mixing 2 primary antibodies for co-staining, they need to be distinguishable by the secondary antibodies. E.g. by species or isotype. Second, possible that the secondary antibody fluorophores are incompatible… e.g. spillover from a fluor in one channel enhancing signal in another channel (though usually this means an increase in signal, not a decrease). Abcam compensation discussion: https://www.abcam.com/index.html?pageconfig=resource&rid=13433 Most literal interpretation but far out on a limb…IL-1Ab is masking the CD11 Ab epitope (either by real biology or antibody background). Not much to do about this but to try alternative antibodies. I know that there is nothing concrete here, sorry. Please let me know if there is anything else I can help you with.

Read More

Answer

I have been in contact with the lab and they have not tested this antibody in flow cytometry, however previous lots have been reported to function for this application. I am trying to find the publications that used this antibody but cannot find them. I am still a little confused as to why when you use the antibody on its own it works perfectly well in flow cytometry but when you try and use CD11 as well, the ab2105 blocks/interferes with the CD11 signal. As you mentioned on the telephone it could be that the blocking reagents used are the issue. I am still trying to consult with my colleagues about this and I will get back to you when I have found out some more information.  

Read More

Answer

Merci pour ces précisions. Comme convenu j'ai procédé à l'envoi d'une unité de ab82558. Le numéro de commande de remplacement gratuit de ce produit est *********. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition. N'hésitez pas à nous contacter lors d'une prochaine occasion.  

Read More

Answer

Suite à votre appel, j'ai regardé les différents anti-IL1 que nous avons dans notre catalogue. Après élimination des anticorps testés avec les protéines recombinantes et non avec des lysats, j'ai retenu les références suivantes : - ab82558 : bande observée à 17 kDa avec cellules Jurkat et MOLT4, mais pas à ~35 kDa. www.abcam.com/ab82558 - ab34837 : www.abcam.com/ab34837, pas d'image mais de bonnes informations concernant la spécificité : "This antibody primarily recognizes the 17kDa mature form of IL1 beta. It also recognizes 10% of the precursor, 31kDa form of IL1 beta in cell lysates. The antibody does not react with IL1 alpha". Comme convenu, je serais ravi de vous envoyer une unité d'un des 2 anticorps mentionnés ci-dessus. Il serait intéressant, je pense, de tester le ab2105 sur cellules Jurkat et MOLT4. Merci de m'indiquer quel anticorps vous souhaitez tester.

Read More

Answer

Thank you for your email! The best way to figure out dilutions and number of tests is to look at the datasheet. In most cases we will have a recommended dilution for each application, and will also include the concentration of the product along with the amount of product in the vial. Using these pieces of information, you can get the number of experiments that you will be able to do with that product. In order to give you the number of tests, I will need to know what application you are looking to use the antibodies in. The calculator at the following link may help you: https://www.msu.edu/~nixonjos/more/dilution.html?stock=1&needed=0.0001&volume=1&total=0.1 To get the number of tests, you can divide the final result ("total stock solution to use") by the amount included in the tube. I hope this helps. If you have any further questions, please do not hesitat

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Shoulder)
Antigen retrieval step
None
Permeabilization
No
Specification
Shoulder
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Paraformaldehyde

Ms. Mindy Cote

Verified customer

Submitted Aug 23 2011

Application
Western blot
Sample
Human Cell lysate - nuclear (keratinocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
keratinocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 26 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human keratinocytes)
Permeabilization
Yes - Triton
Specification
human keratinocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 31 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Keratinocytes)
Permeabilization
Yes - 0,1% Triton-X-100
Specification
Keratinocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 21 2010

21-30 of 38 Abreviews or Q&A

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