Recombinant Anti-IL-18 antibody [EPR19954] (ab207324)
![Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-386638-il18-256952-207324-full--anti-il-18-antibody-epr19954-western-blot--human-lysate.jpg "Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19954] to IL-18
- Suitable for: WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-18 antibody [EPR19954]
See all IL-18 primary antibodies -
Description
Rabbit monoclonal [EPR19954] to IL-18 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra)more 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: Human IL-18 active protein; HeLa, A431, Jurkat, HEK-293, PC-3 and HaCaT whole cell lysates; Human skin and ovary cancer lysates. Flow Cyt (intra): PC-3 and HaCaT 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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19954 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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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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KO cell pellets
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab207324 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 |
1/200. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
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| Flow Cyt (Intra) |
1/60.
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| Notes |
|---|
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WB
1/200. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). |
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Flow Cyt (Intra)
1/60. |
Target
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Function
Augments natural killer cell activity in spleen cells and stimulates interferon gamma production in T-helper type I cells. -
Sequence similarities
Belongs to the IL-1 family. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 3606 Human
- Omim: 600953 Human
- SwissProt: Q14116 Human
- Unigene: 83077 Human
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Alternative names
- Iboctadekin antibody
- IFN gamma inducing factor antibody
- IFN-gamma-inducing factor antibody
see all
Images
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Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IL18 knockout HeLa cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 22 kDaLanes 1-4: Merged signal (red and green). Green - ab207324 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.
ab207324 Anti-IL-18 antibody [EPR19954] was shown to specifically react with IL-18 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265274 (knockout cell lysate ab256952) was used. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. ab207324 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 200 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-334423-anti-il-18-antibody-epr19954-western-blot.jpg)
Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : IL-18 knockout HEK-293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 22 kDaLanes 1 - 2: Merged signal (red and green). Green - ab207324 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab207324 was shown to recognize IL-18 in wild-type HEK293 cells as signal was lost at the expected MW in IL-18 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. Ab207324 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/200 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266047-anti-il-18-antibody-epr19954-western-blot.jpg)
Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)Anti-IL-18 antibody [EPR19954] (ab207324) at 1/1000 dilution + Human IL-18 active protein at 0.02 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266046-anti-il-18-antibody-epr19954-western-blot.jpg)
Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/1000 dilution
Lane 1 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 2 : HaCaT (Human keratinocyte cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The WB profiles are consistent with the literature (PMID 12918059; PMID 11470273).
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![Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266045-anti-il-18-antibody-epr19954-western-blot.jpg)
Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution
Lane 1 : Human skin lysate
Lane 2 : Human ovary cancer lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/5000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Detection in skin and ovary required high concentration of antibody.
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![Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266044-anti-il-18-antibody-epr19954-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling IL-18with ab207324 at 1/60 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
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![Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266043-anti-il-18-antibody-epr19954-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HaCaT (Human keratinocyte cell line) cells labeling IL-18with ab207324 at 1/60 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
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![Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-266042-anti-il-18-antibody-epr19954-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IL-18with ab207324 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
Negative control:
Jurkat cells serve as a negative cell line as described in PMID 15086390.
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![Anti-IL-18 antibody [EPR19954] (ab207324)](https://www.abcam.com/ps/products/207/ab207324/Images/ab207324-9-benefits-of-recombinant-antibodies.png)
Anti-IL-18 antibody [EPR19954] (ab207324)
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (4)
ab207324 has been referenced in 4 publications.
- Zhu H et al. Janus Kinase Inhibition Ameliorates Ischemic Stroke Injury and Neuroinflammation Through Reducing NLRP3 Inflammasome Activation via JAK2/STAT3 Pathway Inhibition. Front Immunol 12:714943 (2021). PubMed: 34367186
- Ye Y et al. Meisoindigo Protects Against Focal Cerebral Ischemia-Reperfusion Injury by Inhibiting NLRP3 Inflammasome Activation and Regulating Microglia/Macrophage Polarization via TLR4/NF-?B Signaling Pathway. Front Cell Neurosci 13:553 (2019). PubMed: 31920554
- Clement M et al. MARK4 (Microtubule Affinity-Regulating Kinase 4)-Dependent Inflammasome Activation Promotes Atherosclerosis-Brief Report. Arterioscler Thromb Vasc Biol 39:1645-1651 (2019). PubMed: 31167564
- Ma ZZ et al. Expression and clinical significance of the NEK7-NLRP3 inflammasome signaling pathway in patients with systemic lupus erythematosus. J Inflamm (Lond) 15:16 (2018). PubMed: 30202244
