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Rabbit Recombinant Monoclonal IL-1 beta antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), WB and reacts with Mouse, Rat, Human samples.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (AB282021), expandable thumbnail
  • Western blot - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (AB282021), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (AB282021), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (AB282021), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (AB282021), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPFlow Cyt (Intra)WBIHC-P
Human
Not recommended
Not recommended
Tested
Tested
Not recommended
Mouse
Tested
Not recommended
Tested
Tested
Not recommended
Rat
Not recommended
Not recommended
Tested
Tested
Not recommended

Tested
Tested

Species
Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Rat, Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Target data

Function

Potent pro-inflammatory cytokine (PubMed:10653850, PubMed:12794819, PubMed:28331908, PubMed:3920526). 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 (PubMed:3920526). Promotes Th17 differentiation of T-cells. Synergizes with IL12/interleukin-12 to induce IFNG synthesis from T-helper 1 (Th1) cells (PubMed:10653850). Plays a role in angiogenesis by inducing VEGF production synergistically with TNF and IL6 (PubMed:12794819). Involved in transduction of inflammation downstream of pyroptosis: its mature form is specifically released in the extracellular milieu by passing through the gasdermin-D (GSDMD) pore (PubMed:33377178, PubMed:33883744). Acts as a sensor of S.pyogenes infection in skin: cleaved and activated by pyogenes SpeB protease, leading to an inflammatory response that prevents bacterial growth during invasive skin infection (PubMed:28331908).

Alternative names

IL1F2, IL1B, Interleukin-1 beta, IL-1 beta, Catabolin

Recommended products

Rabbit Recombinant Monoclonal IL-1 beta antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), WB and reacts with Mouse, Rat, Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR23851-127
Purification technique
Affinity purification Protein A
Specificity

IL-1 beta is not present under homeostatic conditions, and it is induced and secreted only upon inflammatory signals. Stimulation may be required for the detection of IL-1β, as it is not constitutively expressed, no matter in precursor or mature form. For example, when using ab254360, positive signal can be detected in RAW264.7 with 100 ng/ml LPS treatment for 7h and additional 300 ng/ml Brefeldin A treatment in the last 3h. IL-1β is a secreted protein, it is recommended to treat cells with anti-secretion reagents (eg. Brefeldin A) that inhibit secretion.

FURTHER INFORMATION ON SPECIFICITY (Chinese Version) available under the support & downloads section.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Notes

ab282021 is the carrier-free version of Anti-IL-1 beta antibody [EPR23851-127] ab254360.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-IL-1 beta antibody [EPR23851-127] ab254360). Anti-IL-1 beta antibody [EPR23851-127] ab254360 staining IL-1 beta in wild-type THP-1 cells (top panel) and IL1B knockout THP-1 cells (Human IL1B knockout THP-1 cell line ab273762) (bottom panel) +/- TPA (80nM overnight) and LPS (100ng/ml, 6h). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 0.4μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

  • Western blot - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Western blot - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 3% NFDM/TBST

    Nitrocellulose membrane was used in this blot.

    Exposure time: 70 seconds

    All lanes: Western blot - Anti-IL-1 beta antibody [EPR23851-127] (Anti-IL-1 beta antibody [EPR23851-127] ab254360) at 1/1000 dilution

    Lane 1: Untreated NR8383 (rat lung macrophage (alveolar)) whole cell lysate at 20 µg

    Lane 2: NR8383 treated with 100 ng/ml LPS for 7 hours and 300 ng/ml Brefeldin A (Brefeldin A Solution (1,000X) ab193369) for the last 3 hours, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 30 kDa

    Observed band size: 17.5 kDa, 28 kDa, 31 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling IL-1 beta with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 1/50 (10.28 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing increased cytoplasmic and nuclear staining in RAW 264.7 cells treated with Lipopolysaccharide (100 ng/ml) for 4 h then together with Brefeldin A (1 ug/ml) for another 3 h is observed.

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1(Human monocytic leukemia monocyte) treated with 80nM TPA for 16 hours, then 100ng/ml LPS for 3 hours and 300ng/ml BFA for another 3 hours (Right)/ Untreated control (Left) cells labelling IL-1 beta with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 1/500 dilution (0.1ug). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 4 hours and 1ug/ml BFA for another 3 hours (Right)/ Untreated control (Left) cells labelling IL-1 beta with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 1/500 dilution (0.1ug). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NR8383 (rat lung macrophage (alveolar)) treated with 100ng/ml LPS for 4 hours and 1ug/ml BFA for another 3 hours (Right)/ Untreated control (Left) cells labelling IL-1 beta with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 1/50 dilution (1ug)/ Right and Left. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-IL-1 beta antibody [EPR23851-127] ab254360).

    Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with 1 µg/mL LPS and 1 µg/mL BrefeldinA for 18 Hours (bottom), with Anti-IL-1 beta antibody [EPR23851-127] ab254360 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 mins on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody Anti-IL-1 beta antibody [EPR23851-127] ab254360 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 0.2 μg/ml (1/2,500 dilution)) for 30 mins at 22°C . The cells were simultaneously stained with CD14.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins at 22°C

    Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on CD14+.

  • Western blot - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021), expandable thumbnail

    Western blot - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free (ab282021)

    This data was developed using Anti-IL-1 beta antibody [EPR23851-127] ab254360, the same clone in a different buffer formulation.

    Expression of IL-1 beta is induced by LPS treatment. 31-kDa precursor IL-1 beta, 28- and 17.5-kDa proteolytically cleaved IL-1 beta are observed. The expresssion pattern and molecular weight observed is consistent with what has been described in the literature (PMID: 8446594, 19559631).

    Compared with Anti-IL-1 beta antibody [EPR23851-127] ab254360, Anti-IL-1 beta antibody [RM1009] ab283818 has higher affinity. For better using Anti-IL-1 beta antibody [EPR23851-127] ab254360, we recommend loading higher amount of human/rat lysate or using lower antibody dilution.

    Lanes 1 - 6: Western blot - Anti-IL-1 beta antibody [EPR23851-127] (Anti-IL-1 beta antibody [EPR23851-127] ab254360) at 1/1000 dilution

    Lanes 7 - 12: Western blot - Anti-IL-1 beta antibody [RM1009] (Anti-IL-1 beta antibody [RM1009] ab283818)

    Lanes 1 and 7: Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lanes 2 and 8: THP-1 treated with 100ng/ml lipopolysaccharide (LPS) for 3h whole cell lysate at 20 µg

    Lanes 3 and 9: Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lanes 4 and 10: RAW 264.7 treated with 100 ng/ml LPS and 300 ng/ml Brefeldin A (Brefeldin A Solution (1,000X) ab193369) for 4 hours, whole cell lysate at 20 µg

    Lanes 5 and 11: Untreated NR8383 (rat lung macrophage (alveolar)) whole cell lysate at 20 µg

    Lanes 6 and 12: NR8383 treated with 100 ng/ml LPS for 7 hours and 1 µg/ml Brefeldin A (Brefeldin A Solution (1,000X) ab193369) for the last 3 hours, whole cell lysate at 20 µg

    Observed band size: 17.5,28,31 kDa

    Exposure time: 102s

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Product protocols

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