Overview

  • Product name
  • Description
    Rabbit polyclonal to IL-1 beta
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ELISA, Neutralising, IHC-P, ICC, IHC-Fr, IHC-FoFr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Full length protein aa 118-269. Full length mature protein minus the propeptide from aa 1-117.
    Database link: P10749

  • Positive control
    • Recombinant mouse IL-1 beta protein (ab9723) can be used as a positive control in WB. FFPE mouse kidney tissue sections
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab9722 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 dilution. To detect mIL-1b by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant mIL-1b is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.
ELISA Use at an assay dependent dilution. To detect mIL-1b by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant mIL-1b.
Neutralising Use at an assay dependent dilution. To yield one-half maximal inhibition [ND50] of the biological activity of mIL-1b (50 pg/ml), a concentration of 100 - 150 ng/ml of this antibody is required.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC 1/100.
IHC-Fr Use at an assay dependent dilution. PubMed: 18420712Acetone fixed.
IHC-FoFr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

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

  • ab9722 staining IL-1 beta in murine bone marrow-derived macrophages by Immunocytochemistry/ Immunofluorescence. Cells are immortalised murine bone marrow-derived macrophages stably transfected with GFP-LC3 (green) to visualise autophagosomes. The cells were fixed in paraformaldehyde, permeabilised in 0.01% Triton X-100 and then blocked using 5% serum for 1 hour at 20°C. Samples were then incubated with primary antibody at 1/200 for 1 hour at 20°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 568 (red) used undiluted.

    See Abreview

  • ab9722 staining IL-1 beta in rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in diluent) for 10 hours at 25°C. An AlexaFluor®555-conjugated donkey anti-goat IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • ab9722 staining IL-1 beta (green) in murine macrophage cells by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 + 3% BSA and blocked with 3% BSA for 3 hours at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 1 hour at 25°C. A FITC-conjugated goat anti-rabbit polyclonal IgG (1/100) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

  • IHC image of IL-1 beta staining in mouse kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9722, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemical analysis of PFA perfusion fixed frozen mouse brain, staining IL-1 beta (green) with ab9722 at 0.5 &microg/ml. A peroxidase conjugated secondary antibody was used and staining was detected using a fluorescein Tyramide Signal Amplification (TSA™) reagent.

  • Sandwich ELISA detecting IL-1 beta using ab9722 at a concentration of 0.1 µg/ml.

References

This product has been referenced in:
  • Chauhan G  et al. Distinct influence of COX-1 and COX-2 on neuroinflammatory response and associated cognitive deficits during high altitude hypoxia. Neuropharmacology 146:138-148 (2019). Read more (PubMed: 30476507) »
  • Aboutaleb N  et al. Lavender oil (Lavandula angustifolia) attenuates renal ischemia/reperfusion injury in rats through suppression of inflammation, oxidative stress and apoptosis. Biomed Pharmacother 110:9-19 (2019). Read more (PubMed: 30453254) »
See all 201 Publications for this product

Customer reviews and Q&As

1-10 of 13 Q&A

Question
Answer

The Il-1 beta antibody ab9722 is tested for reactivity against recombinant mature active IL-1 beta. We do not have any in-house data demonstrating reactivity with the precursur/pro-form, but we at least predict reactivity with the denatured pro-form, given that it contains the amino acid sequence of the active form. One review from a customer seems to demonstrate reactivity with the pro-form in a western blot of lysates of macrophages +/- LPS stimulation. Here is a link to the review.

https://www.abcam.com/IL1-beta-antibody-ab9722/reviews/9574

However, the antibody will not be able to distinguish the pro-form from the mature form in other immunoassays such as immunohistochemistry or ELISA.

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Answer

Thank you for contacting us. There is no MSDS for ab9722, as it contains only PBS and no hazardous chemicals.

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Answer

Thank you for your enquiry.

As far as we are aware, ab9722 IL1 beta antibodyhas been tested and guaranteed in mouse but nottested in rat. the other ab9787IL1 beta antibody is tested and guaranteed for rat but not tested in mouse.

As requested, I can confirm the following alignments:

ab9722 Anti-IL1 beta antibody
Already tested mouse.Alignment to rat: 89%

ab9787 Anti-IL1 beta antibody
Already tested rat.Alignment to mouse: 90%

These alignments are high, so theoretically it is likely that ab9722 will detect in rat and that ab9787 will detect in mouse which I hope is good news for your customer. However, please note we are not able to guarantee this reactivity without further testing. If the customerwould like to test the antibody in this untested species, please contact me again by replying to this message prior to the purchase asthey may be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

Read More

Answer

Thank you for your reply, and I apologize that I didn't properly caption the image that I sent.

The Western blot was run on murine recombinant mature IL1 beta, not on cell lysates. The lab does not test this antibody with cell lysates.

The lanes represent different loading amounts of the recombinant protein: 250, 125, 62.5, 31.25, 15.625, 7.8, 3.9, 1.95, 0.975, 0.4875, and 0.24 ng/lane left to right. The antibody was used at 0.2 ug/mL.

We have not done in-house testing on the pro IL1 beta, but customers have reported seeing a higher molecular weight band that we believe is the precursor protein.

I apologize again that I did not previously send this information. Please let me know if you have any further questions about this.

Read More

Answer

Thank you for your call last week and for your patience while I have been in touch with the lab.

The lab tests this antibody with recombinant murine IL1 beta, and a strong band around 17 kDa is seen in addition to a band around 34 kDa that is believed to be the dimer. I have attached the QC results for the lot of ab9722 that you have most recently used.

At this time we are not sure what is causing the bands around 38 and 54 kDa with the treated RAW cell lysates and tissue lysates, and I've looked through the literature but have found no other references to these band sizes. We don't have another unconjugated IL1 beta antibody that is known to react with mouse samples, but I would be happy to send a replacement antibody of your choice or issue a credit or refund if you prefer. We do have an HRP-conjugatedanti-mouse IL1 beta antibody, ab106035, which you might be interested to try:

https://www.abcam.com/IL1-beta-antibody-HRP-ab106035.html

I am sorry that I don't have more information this time, but please let me know if you are interested in receiving a different antibody or if you have any further questions. I look forward to hearing from you.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab106035.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thank you for sending these answers and the images. They are very helpful to better understand the situation.

Were the tissue sections taken from diseased or treated brain? I'm not an expert on IL1 beta, but it seems that it is highest expressed in inflammed tissue. If that is the case, then I would probably expect positive cells to be fewer and more spread out than the appear in the images. Do you have access to any tissue that would be less than a year old, just to see if the tissue is damaged which has caused non-specific binding? Or you could try omitting the primary antibody and just using the secondary antibody, to see if antibodies in general are sticking to the tissue.

We do guarantee all of our products to work as stated on the datasheet for up to 6 months after purchase, so I would be happy to send a replacement or alternatively issue a credit or refund if you would like. Ab9722 is the antibody Il2 beta antibody that we have that has been tested in mouse frozen tissue staining, however I could send a different lot if we have one in stock. Do you know your original order or PO number?

I look forward to hearing from you and resolving this promptly. Please let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for contacting us and letting us know about the trouble with this antibody.

Would you mind if I ask a few questions about the results and protocol, in order to see if there is anything that I might suggest to improve the results?

1) Is there any staining at all (even non-specific) on the tissue, or is thereno staining?Could you provide an image?
2) What species is the brain sample from?
3) How was the brain tissue fixed and prepared for staiing(e.g. antigen retrieval, permeabilization, etc)?
4) What dilution of antibody was used?
5) What kind of blocking agent was used (e.g. serum or BSA)?
6)What kind of secondary antibody and detection system were used?

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

Answer

Thank you for your reply. It is likely that ab106034 would meet your requirements; we do not currently have any images for this antibody though. It is covered under our Abpromise though to work on mouse and rat tissue in IHC-P and western blotting. As we do not currently have an image of ab106034 in IHC-P, I can offer a discount off a future purchase if you buy ab106034 now, use it  IHC-P and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free PRIMARY ANTIBODY. It doesn’t matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and send an image of ab106034  in IHC-P. I will then send a discount code. This code must be issued before purchasing ab106034  so please wait for my reply before ordering. 2. Purchase ab106034 either by phone, fax, or online (www.abcam.com). 3. Use it in IHC-P. 4. Send us an image of the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODYordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.    

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Answer

I have been searching through our catalogue and I still think the best antibody for you would be ab9722. If there is anything else I can help you with, please let me know.

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1-10 of 13 Q&A

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