Anti-IL-1 beta antibody [EPR23851-127] 20 ul size (ab254360)

Anti-IL-1 beta antibody [EPR23851-127] is a rabbit recombinant monoclonal antibody that is used to detect IL-1 beta in Flow cytometry (Intra), ICC/IF, Western blot. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with Il1b knockout cell line validation
- Stimulation may be required for detection of Il-1 beta
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 90 publications

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Images

Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] (AB254360), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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/IF	IP	WB	IHC-P	Flow Cyt (Intra)
Human	Not recommended	Not recommended	Tested	Not recommended	Tested
Mouse	Tested	Not recommended	Tested	Not recommended	Tested
Rat	Not recommended	Not recommended	Tested	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat, Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2500	Notes -
Species Human	Dilution info 1/2500	Notes -
Species Rat	Dilution info 1/2500	Notes -

Target data

See full target information IL1B

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

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

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

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-IL-1 beta antibody [EPR23851-127] (ab254360) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF and WB.

Anti-IL-1 beta antibody [EPR23851-127] (ab254360) was first used in a scientific publication in 2021 and has been cited over 96 times in peer reviewed journals. It's performance in Western blot in human, mouse and rat samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-IL-1 beta antibody [EPR23851-127] (ab254360) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-IL-1 beta antibody [EPR23851-127] (ab254360) has been confirmed by ICC/IF testing in IL-1 beta knockout THP-1 cells (Human IL1B knockout THP-1 cell line ab273762).

Anti-IL-1 beta antibody [EPR23851-127] (ab254360) specifically detects IL-1 beta (UniProt ID: P10749; Molecular weight: 18kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Conjugation-ready, carrier free format available for antibody clone EPR23851-127 - Anti-IL-1 beta antibody [EPR23851-127] - BSA and Azide free ab282021.

Antibody clone EPR23851-127 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, Alexa Fluor^® 647 (Alexa Fluor® 488 Anti-IL-1 beta antibody [EPR23851-127] ab317662, Alexa Fluor® 647 Anti-IL-1 beta antibody [EPR23851-127] ab319085).

Interleukin-1 beta (IL-1β) is a key biomarker in inflammatory bowel disease (IBD).

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

What are the advantages of a recombinant monoclonal antibody?
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.

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

Flow Cytometry (Intracellular) - Anti-IL-1 beta antibody [EPR23851-127] (ab254360)
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 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.
Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR23851-127] (ab254360)
ab254360 staining IL-1 beta in wild-type THP-1 cells and IL1B knockout THP-1 cells (Human IL1B knockout THP-1 cell line ab273762) +/- 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 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] (ab254360)
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] (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] (ab254360)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling IL-1 beta with ab254360 at 1/50 (10.28 μg/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 μg/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] (ab254360)
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 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] (ab254360)
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 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.
Western blot - Anti-IL-1 beta antibody [EPR23851-127] (ab254360)
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 ab254360, Anti-IL-1 beta antibody [RM1009] ab283818 has higher affinity. For better using 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] (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
Flow Cytometry (Intracellular) - 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 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 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+.