Overview

  • Product name
  • Description
    Rabbit polyclonal to IL1 beta
  • 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
    • FFPE mouse kidney tissue sections

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 IL1 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 IL1 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 IL1 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 IL1 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 IL1 beta (green) with ab9722 at 0.5 µg/ml. A peroxidase conjugated secondary antibody was used and staining was detected using a fluorescein Tyramide Signal Amplification (TSA™) reagent.

References

This product has been referenced in:
  • Wu J  et al. Roles of programmed death protein 1/programmed death-ligand 1 in secondary brain injury after intracerebral hemorrhage in rats: selective modulation of microglia polarization to anti-inflammatory phenotype. J Neuroinflammation 14:36 (2017). WB ; Rat . Read more (PubMed: 28196545) »
  • Zhu Y  et al. Electro-acupuncture promotes the proliferation of neural stem cells and the survival of neurons by downregulating miR-449a in rat with spinal cord injury. EXCLI J 16:363-374 (2017). Read more (PubMed: 28507480) »

See all 85 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Oct 30 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (fat and muscle)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
fat and muscle
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Oct 30 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
tumor xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Oct 30 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
skin tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Oct 30 2017

Application
Flow Cytometry
Sample
Rat Cell (Brain)
Permeabilization
Yes - 0.25% Triton X-100
Gating Strategy
single cells population
Specification
Brain
Fixation
Paraformaldehyde
Username

Ms. Aneta Radziwon-Balicka

Verified customer

Submitted Sep 12 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (spinal cord)
Permeabilization
Yes - 0.1%triton+0.6%H2O2 in 1XTBST
Specification
spinal cord
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 08 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Antigen retrieval step
None
Sample
Rat Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Paraformaldehyde
Username

Brandon Jun

Verified customer

Submitted Dec 30 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (hepatocytes)
Specification
hepatocytes
Treatment
20 ug/ml poly(I:C)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Dec 17 2014

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, g...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW264.7)
Specification
RAW264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton + 2% BSA in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Apr 12 2013

1-10 of 31 Abreviews or Q&A

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