Recombinant Anti-CXCL9 antibody [EPR23999-5] (ab263442)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23999-5] to CXCL9
- Suitable for: Flow Cyt (Intra), WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CXCL9 antibody [EPR23999-5]
See all CXCL9 primary antibodies -
Description
Rabbit monoclonal [EPR23999-5] to CXCL9 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IPmore details
Unsuitable for: ICC/IF or IHC-P -
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: THP-1 (treated with 200ng/ml IFN-? and 50 ng/ml LPS(Lipopolysaccharide) for 24 hours) whole cell lysate; His-tagged human purified recombinant full-length protein CXCL9. Flow Cyt (intra): THP-1 (treated with 100ng/ml IFN-gamma and 1ug/ml LPS(Lipopolysaccharide) for 48 hours) cells. IP: THP-1 (treated with 100ng/ml IFN-gamma and 1ug/ml LPS(Lipopolysaccharide) for 48 hours) whole cell lysate.
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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
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23999-5 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Coimmunogen
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab263442 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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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 13, 14 kDa (predicted molecular weight: 14 kDa).
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 13, 14 kDa (predicted molecular weight: 14 kDa). |
IP
1/30. |
Target
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Function
Cytokine that affects the growth, movement, or activation state of cells that participate in immune and inflammatory response. Chemotactic for activated T-cells. Binds to CXCR3. -
Sequence similarities
Belongs to the intercrine alpha (chemokine CxC) family. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 4283 Human
- Omim: 601704 Human
- SwissProt: Q07325 Human
- Unigene: 77367 Human
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Alternative names
- C-X-C motif chemokine 9 antibody
- chemokine (C-X-C motif) ligand 9 antibody
- CMK antibody
see all
Images
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All lanes : Anti-CXCL9 antibody [EPR23999-5] (ab263442) at 1/1000 dilution
Lane 1 : Wild-type THP-1 untreated control cell lysate
Lane 2 : Wild-type THP-1 treated IFNg (200 ng/mL, 24 h) + LPS (50 ng/mL, 24 h) cell lysate
Lane 3 : CXCL9 knockout THP-1 untreated control cell lysate
Lane 4 : CXCL9 knockout THP-1 treated IFNg (200 ng/mL, 24 h) + LPS (50 ng/mL, 24 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CXCL9 antibody [EPR23999-5] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab263442 was shown to bind specifically to CXCL9. A band was observed at 17 kDa in treated wild-type THP-1 cell lysates with no signal observed at this size in CXCL9 knockout cell line ab282347 (knockout cell lysate ab283019). To generate this image, wild-type and CXCL9 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-CXCL9 antibody [EPR23999-5] (ab263442) at 1/1000 dilution
Lane 1 : Untreated THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 2 : THP-1 (treated with 200 ng /ml IFN-gamma and 50 ng/ml LPS (Lipopolysaccharide) for 24 hours), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 14 kDa
Observed band size: 13,14 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile is consistent with what has been described in the literature (PMID: 15154616, 22973274).
Exposure time: 3 minutes.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized THP-1 (human monocytic leukemia monocyte) treated with 100ng/ml IFN-gamma and 1ug/ml LPS (Lipopolysaccharide) for 48 hours (Red)/ Untreated control (Green) cells labelling CXCL9 with ab263442 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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CXCL9 was immunoprecipitated from 0.35 mg THP-1(human monocytic leukemia monocyte) (treated with 100ng/ml IFN-γ and 1ug/ml LPS (Lipopolysaccharide) for 48 hours) whole cell lysate with ab263442 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263442 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 treated with 100ng/ml IFN-γ and 1ug/ml LPS (Lipopolysaccharide) for 48 hours, whole cell lysate 10 ug
Lane 2: ab263442 IP in THP-1 treated with 100ng/ml IFN-γ and 1ug/ml LPS(Lipopolysaccharide) for 48 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263442 in THP-1 treated with 100ng/ml IFN-γ and 1ug/ml LPS(Lipopolysaccharide) for 48 hours whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Anti-CXCL9 antibody [EPR23999-5] (ab263442) at 1/1000 dilution + His-tagged human purified recombinant full-length protein CXCL9, 10 ng
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 14 kDa
Observed band size: 13,14 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile is consistent with what has been described in the literature (PMID: 15154616, 22973274).
Exposure time: 5.5 seconds.
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
Certificate of Compliance
References (0)
ab263442 has not yet been referenced specifically in any publications.