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

11-20 of 38 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (peripheral blood mononuclear cells)
Gel Running Conditions
Reduced Non-Denaturing (Native) (5-16% gel)
Loading amount
25 µg
Treatment
see below
Specification
peripheral blood mononuclear cells
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 07 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1 human leukemic monocyte)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
100 nM PMA for 24 hrs, 100 ng/ml LPS for 4 hrs
Specification
THP-1 human leukemic monocyte
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Aug 12 2013

Answer


We have 3 IL-1B antibodies that have been tested in IHC-P and WB with human samples and that as well state to recognize the active/mature form:

ab2105, ab34837, ab106015
https://www.abcam.com/il1-beta-antibody-ab2105.html
https://www.abcam.com/il1-beta-antibody-ab34837.html
https://www.abcam.com/il1-beta-antibody-hrp-ab106015.html


However, please keep in mind since the mature form (17 kDa) is part of the unprocessed form (31 kDa), that especially in WB, any antibody binding to the active form will also bind to the unprocessed form as the active sequence is contained in the unprocessed form.

The datasheets for these antibodies state that the antibodies will recognize 10% of the unprocessed protein, especially in WB.

Read More

Answer

Vielen Dank für Ihre Anfrage die Herr Hryca uns in den technischen Dienst weitergeleitet hat.

Ich habe geschaut, und wir haben viele möglichen positiven Kontrollen für Sie. Können Sie mir bitte noch sagen, welche Art von positive Kontrolle Sie benötigen? Ist es ein Lysat für Western blot oder ein Gewebeschnitt für Immunohistochemie? Haben Sie auch eine Präferenz für eine Spezies (Maus, human)?

Hier mal eine Aufstellung von möglichen positiven Kontrollen. Ich freue mich wieder von Ihnen zu hören, um Ihnen genaure Vorschläge machen zu können.




cell lines Slide
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=2105 Rabbit polyclonal to IL1 beta LPS treated RAW 264.7 cells, Alveolar macrophages (MHS cell line) liver, diseased, inflammed tissue, monocytes, macrophages
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6313 Mouse monoclonal [CTD-19] to Cathepsin D SKBR3, MCF7 Human liver tissue.
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6535 Mouse monoclonal [BRM-22] to Hsp70 Hela, 293T,MCF7, Jurkat, HepG2, A431, HCT116, PC3, ... liver, brain, testes, heart, skeletal muscle
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=8219 Mouse monoclonal [MEM-259] to CD63 HT1080, HepG2, OVCAR-3, HL60 and A375 hemapoietic ells, tissue macrophages, platelet, human melanoma tissue, monocytes, macrophages, endothelial cells, SM cells, brain white matter, peripheral nerves
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=25631 Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker HepG2, 3T3, HeLa, RAW264.7, ... placenta, lung, liver
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=50533 Mouse monoclonal [Rab7-117] to RAB7 HepG2, B16, SK-MEL-28, 3T3, Human brain whole tissue, mouse neuroblastoma
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=68340 Mouse monoclonal to Rab9 Hela, A172, 3T3, 293, PC-12. COS-7, THP-1 tonsil, spleen, prostate cancer, stomach tissu
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=72907 Rabbit polyclonal to Rab5b Hela, Jurkat, HEK293, HepG2 Human Placenta Tissue, Human Brain Tissue.

Read More

Answer

According to our records, ab2105 was proving difficult to use in WB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Read More

Answer

Thank you for contacting us.

To our knowledge, customised protocols are not required for this product. Any general WB protcol will work.

The dilution 1/100-1/200 are recommended for guidelines only. If you observe high background try the antibody at 1/1000 dilution; overall the working conditions has to determined by the end user.

We use 5% BSA fro blocking in our lab.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for contacting Abcam regarding ab2105.


I am sorry that you have been experiencing difficulties with this antibody in WB.I was wondering if you could provide some additional information regarding your sample - what species, tissue, and method of preparation (lysis buffer, etc)was used? Have you tested other dilutions? What is the blocking buffer and antibody dluent? Can you provide an image of your blot?


Once I have some additional information, it will be easier to determine if modifications to your protocol may improve your results or if the antibody is at fault. Also, please provide a lot number and order number.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

Thank you for contacting Abcam.

Please find attached the requested Certificate of Compliances.

If there is anything else I can help you, please let me know.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Keratinocytes)
Permeabilization
Yes - 0,1% Saponin / PBS
Specification
Keratinocytes
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jan 21 2012

Answer

Thank you for your valuable comments. This antibody is sourced and unfortunately we do not have another Western blot image to upload. We have decided to remove this Western blot image; further optimization may be necessary. We will update datasheet with a new image as soon as there are some available. We apologize for any inconvenience caused and thank you for bringing this matter to our attention.

Read More

11-20 of 38 Abreviews or Q&A

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