Recombinant Anti-IL-1 beta antibody [RM1009] (ab283818)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit recombinant multiclonal [RM1009] to IL-1 beta
- Suitable for: IHC-P, WB, ICC, IP, Flow Cyt
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-1 beta antibody [RM1009]
See all IL-1 beta primary antibodies -
Description
Rabbit recombinant multiclonal [RM1009] to IL-1 beta -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC, IP, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
This product was produced with the following immunogens:
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers. -
Positive control
- WB: U-87 MG ; PMA, LPS & BFA treated U-937 ; LPS & BFA treated RAW 264.7 & NR8383 whole cell lysates. IHC-P: Mouse colon carcinoma tissue ; LPS & BFA treated Mouse lung tissue & RAW 264.7. ICC: PMA, LPS & BFA treated U-937 cells ; LPS & BFA treated J774A.1 & RAW 264.7 cells. Flow cyt: PMA, LPS & BFA treated U-937 cells ; LPS & BFA treated J774A.1 cells. IP: PMA, LPS & BFA treated U-937 cells ; LPS & BFA treated J774A.1 & RAW 264.7 cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Clonality
Recombinant Multiclonal -
Clone number
RM1009 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283818 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 |
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
1/1000.
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ICC |
1/50.
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IP |
1/30.
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Flow Cyt |
1/500.
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Notes |
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IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000. |
ICC
1/50. |
IP
1/30. |
Flow Cyt
1/500. |
Target
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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
modificationsActivation 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
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Database links
- Entrez Gene: 3553 Human
- Entrez Gene: 16176 Mouse
- Entrez Gene: 24494 Rat
- Omim: 147720 Human
- SwissProt: P01584 Human
- SwissProt: P10749 Mouse
- SwissProt: Q63264 Rat
- Unigene: 126256 Human
see all -
Alternative names
- Catabolin antibody
- H1 antibody
- IFN beta inducing factor antibody
see all
Images
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All lanes : Anti-IL-1 beta antibody [RM1009] (ab283818) at 1/1000 dilution
Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 2 : U-937 (Human histiocytic lymphoma monocyte) whole cell lysate
Lane 3 : U-937 treated with 100nM PMA for 2 days, then 1µg/ml LPS for 13h and add 5µg/ml BFA for another 4 h whole cell lysate
Lane 4 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 5 : RAW 264.7 treated with 100ng/ml LPS for 4h and add 1µg/ml BFA for another 3 h whole cell lysate
Lane 6 : NR8383 (Rat lung macrophage (alveolar)) whole cell lysate
Lane 7 : NR8383 treated with 100ng/ml LPS for 4h and add 1µg/ml BFA for another 3 h whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Observed band size: 17.5,28,31 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
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 expression pattern and molecular weight observed is consistent with what has been described in the literature (PMID: 8446594, 19559631)
Exposure time: Lane 1-3: 3 min ; Lane 4-7: 70 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 beta antibody [RM1009] (ab283818)
Immunohistochemical analysis of paraffin-embedded Mouse colon carcinom tissue labelling IL-1 beta with ab283818 at 1/500 (1.058 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on the lamina propria in mouse colon carcinoma. The section was incubated with ab283818 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 beta antibody [RM1009] (ab283818)
Immunohistochemical analysis of paraffin-embedded RAW 264.7 cells labelling IL-1 beta with ab283818 at 1/500 (1.058 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on RAW 264.7 cells treated with LPS (100ng/ml for 4+3 hours) added BFA (1µg/ml for 3 hours) (Image A) and no staining on control RAW 264.7 (Image B). The section was incubated with ab283818 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 beta antibody [RM1009] (ab283818)
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labelling IL-1 beta with ab283818 at 1/500 (1.058 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on mouse lung treated with LPS (1ug/ml for 16 hours) and BFA (1ug/ml for 16h hours) (Image A) and no staining on control mouse lung was observed (Image B). The section was incubated with ab283818 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 beta antibody [RM1009] (ab283818)
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling IL-1 beta with ab283818 at 1/500 (1.058 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). The section was incubated with ab283818 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Negative control: no staining on mouse colon.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 beta antibody [RM1009] (ab283818)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling IL-1 beta with ab283818 at 1/500 (1.058 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). The section was incubated with ab283818 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Negative control: no staining on mouse spleen.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-937 cells labelling IL-1 beta with ab283818 at 1/50 (10.58 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in U-937 cells treated with Phorbol-12-myristate-13-acetate (100 nM) for 2 days, then replaced it with Lipopolysaccharides (1 µg/ml) for 13 h, with the addition of Brefeldin A (5 µg/ml) for the last 4 hours. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 cells labelling IL-1 beta with ab283818 at 1/50 (10.58 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in J774A.1 cells treated with Lipopolysaccharides (100 ng/ml) for 4h, with the addition of brefeldin A (1 ug/ml) for the last 3 hours. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling IL-1 beta with ab283818 at 1/50 (10.58 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic and nuclear staining in RAW 264.7 cells transfected with Lipopolysaccharides (100 ng/ml) for 4h, with the addition of brefeldin A (1 ug/ml) for the last 3 hours. (PMID:18490713, PMID:18939951). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-937 (Human histiocytic lymphoma monocyte) treated with 100nM PMA for 2days then 1µg/ml LPS treated for 13h and add 5µg/ml BFA for another 4h (Red) / U-937 treated with 100nM PMA for 2days (Green) cells labelling IL-1 beta with ab283818 at 1/500 dilution (0.1ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) treated with 100ng/ml LPS treated for 4h and add 1µg/ml BFA for another 3h (Red) / Untreated control (Green) cells labelling IL-1 beta with ab283818 at 1/500 dilution (0.1ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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IL-1 beta was immunoprecipitated from 0.35 mg U-937 (Human histiocytic lymphoma monocyte) treated with 100nM PMA for 2 days, then 1µg/ml LPS for 13h and add 5µg/ml BFA for another 4 h, whole cell lysate 10ug with ab283818 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283818 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: U-937 (Human histiocytic lymphoma monocyte) treated with 100nM PMA for 2 days, then 1µg/ml LPS for 13h and add 5µg/ml BFA for another 4 h, whole cell lysate 10ug
Lane 2: ab283818 IP in U-937 treated with 100nM PMA for 2 days, then 1µg/ml LPS for 13h and add 5µg/ml BFA for another 4 h, whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283818 in U-937 treated with 100nM PMA for 2 days, then 1µg/ml LPS for 13h and add 5µg/ml BFA for another 4 h, whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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IL-1 beta was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 4h and add 1ug/ml BFA for another 3 h whole cell lysate 10ug with ab283818 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283818 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 4h and add 1ug/ml BFA for another 3 h whole cell lysate 10ug
Lane 2: ab283818 IP in RAW 264.7 treated with 100ng/ml LPS for 4h and add 1ug/ml BFA for another 3 h whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283818 in RAW 264.7 treated with 100ng/ml LPS for 4h and add 1ug/ml BFA for another 3 h whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab283818 has not yet been referenced specifically in any publications.