Recombinant Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)
![Western blot - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)](https://www.abcam.com/ps/products/229/ab229696/Images/ab229696-296159-anti-il-1-beta-antibody-epr21086-western-blot.jpg "Western blot - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21086] to IL-1 beta - BSA and Azide free
- Suitable for: ICC/IF, WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free
See all IL-1 beta primary antibodies -
Description
Rabbit monoclonal [EPR21086] to IL-1 beta - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: LPS treated THP-1 whole cell lysate. IP: LPS treated THP-1 whole cell lysate. ICC/IF: U-937 cells treated with Phorbol-12-myristate-13-acetate, then LPS and BFA.
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General notes
ab229696 is the carrier-free version of ab216995.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21086 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229696 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 |
|---|---|---|
| ICC/IF |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Detects a band of approximately 35, 29, 17 kDa (predicted molecular weight: 30 kDa).
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| IP |
Use at an assay dependent concentration.
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| Notes |
|---|
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ICC/IF
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Detects a band of approximately 35, 29, 17 kDa (predicted molecular weight: 30 kDa). |
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IP
Use at an assay dependent concentration. |
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
- Omim: 147720 Human
- SwissProt: P01584 Human
- Unigene: 126256 Human
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Alternative names
- Catabolin antibody
- H1 antibody
- IFN beta inducing factor antibody
see all
Images
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Western blot - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)All lanes : Anti-IL-1 beta antibody [EPR21086] (ab216995) at 1/1000 dilution
Lane 1 : Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 2 : THP-1 treated with 100 ng/ml Lipopolysaccharide (LPS) for 3 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 30 kDa
Exposure time: 3 minutesBlocking/Dilution buffer and concentration: 5% NFDM/TBST.
The expression profile is consistent with the literature (PMID 15192144 and 10845914).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216995).
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![Western blot - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)](https://www.abcam.com/ps/products/229/ab229696/Images/ab229696-453267-anti-il-1-beta-antibody-epr21086-western-blot-wildtype-thp1-untreated-wildtype-thp1-lps-treated-3-h-100-ngml-il1b-k.jpg)
Western blot - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)All lanes : Anti-IL-1 beta antibody [EPR21086] (ab216995) at 1/1000 dilution
Lane 1 : Wild-type THP-1 untreated cell lysate
Lane 2 : Wild-type THP-1 LPS-treated (3 h, 100 ng/ml) cell lysate
Lane 3 : IL1B knockout THP-1 untreated cell lysate
Lane 4 : IL1B knockout THP-1 LPS-treated (3 h, 100 ng/ml) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 27-32 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab216995).
Lanes 1 - 4: Merged signal (red and green). Green - ab216995 observed at 27-32 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab216995 was shown to react with IL-1 beta in wild-type THP-1 cells in Western blot with loss of signal observed in IL1B knockout sample. Wild-type THP-1 and IL1B knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab216995 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)](https://www.abcam.com/ps/products/229/ab229696/Images/ab229696-472642-antiil1-beta-antibody-epr21086-immunocytochemistry.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216995).
Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.1% TritonX-100 permeabilised U-937 (human histiocytic lymphoma monocyte) cells labeling IL-1 beta with ab216995 at 1/50 (10 µg/ml) dilution. Followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 µg/ml) dilution. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 µg/ml) dilution. Nuclear staining: DAPI.
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 addition of Brefeldin A (5 µg/ml) for the last 4 hours.
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![Immunoprecipitation - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)](https://www.abcam.com/ps/products/229/ab229696/Images/ab229696-321380-anti-il-1-beta-antibody-epr21086-immunoprecipitation.jpg)
Immunoprecipitation - Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)IL-1 beta was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate treated with 100 ng/ml lipopolysaccharide (LPS) for 3 hours, with ab216995 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216995 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: THP-1 (human monocytic leukemia monocyte) treated with 100 ng/ml lipopolysaccharide (LPS) for 3 hours whole cell lysate 10 µg (Input).
Lane 2: ab216995 IP in THP-1 treated with 100 ng/ml lipopolysaccharide (LPS) for 3 hours whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216995 in THP-1 treated with 100 ng/ml lipopolysaccharide (LPS) for 3 hours whole cell lysate (-).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216995).
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![Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)](https://www.abcam.com/ps/products/229/ab229696/Images/ab229696-3-benefits-of-recombinant-antibodies.png)
Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)
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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Datasheet download
Certificate of Compliance
References (0)
ab229696 has not yet been referenced specifically in any publications.
