Recombinant Anti-IL-18 antibody [EPR19954] KO Tested (ab207324)

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Primary - Rabbit Anti-IL-18 antibody [EPR19954] (ab207324)
Flow Cyt, WB

Protein - Recombinant Human IL-18 protein (ab188059)
SDS-PAGE, HPLC

View more associated products

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: Flow Cyt, WBmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment within Human IL-18 aa 1 to the C-terminus. The exact sequence is proprietary. Corresponds to the mature protein minus the propeptide.
Database link: Q14116
Positive control
- WB: Human IL-18 active protein; PC-3 and HaCaT whole cell lysates; Human skin and ovary cancer lysates. Flow Cyt: PC-3 and HaCaT cells.
General notes

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 rabbit monoclonal antibody.

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

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19954
Isotype

IgG
Research areas

Alternative Versions
- Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
Compatible Secondaries
- Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human IL-18 protein (ab188059)

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
Flow Cyt		1/60.
WB		1/200. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

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.
Target information above from: UniProt accession Q14116 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3606 Human
- Omim: 600953 Human
- SwissProt: Q14116 Human
- Unigene: 83077 Human
Alternative names
- Iboctadekin antibody
- IFN gamma inducing factor antibody
- IFN-gamma-inducing factor antibody
- IGIF antibody
- IL 1 gamma antibody
- IL 18 antibody
- IL 1g antibody
- IL-1 gamma antibody
- IL-18 antibody
- IL1 gamma antibody
- IL18 antibody
- IL18 protein antibody
- IL18_HUMAN antibody
- IL1F4 antibody
- IL1g antibody
- IL1gamma antibody
- ILIF4 antibody
- Interferon gamma inducing factor antibody
- Interferon gamma-inducing factor antibody
- Interleukin 1 gamma antibody
- Interleukin 18 (interferon-gamma-inducing factor) antibody
- Interleukin 18 antibody
- Interleukin-1 gamma antibody
- Interleukin-18 antibody
- Interleukin18 antibody
- MGC12320 antibody
see all

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 kDa

Lanes 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.
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 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
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 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The WB profiles are consistent with the literature (PMID 12918059; PMID 11470273).
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 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Detection in skin and ovary required high concentration of antibody.
Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

Flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling IL-18 with 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 Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

Flow cytometric analysis of 4% paraformaldehyde-fixed HaCaT (Human keratinocyte cell line) cells labeling IL-18 with 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 Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IL-18 with 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 Fluor^® 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.

Western blot protocols
Flow cytometry protocols

Datasheet

This product has been referenced in:

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). Read more (PubMed: 30202244) »

See 1 Publication for this product

Publishing research using ab207324? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Recombinant Anti-IL-18 antibody [EPR19954] (ab207324)

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References

Customer reviews and Q&As