Recombinant Anti-IL-8 antibody [EPR22994-255] (ab235584)
![Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)](https://www.abcam.com/ps/products/235/ab235584/Images/ab235584-453256-anti-il-8-antibody-epr22994-255-western-blot-wildtype-pc3-brefeldin-a-5-ugml-5-h-wildtype-pc3-lps-2-ugml-6-h-with.jpg "Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22994-255] to IL-8
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-8 antibody [EPR22994-255]
See all IL-8 primary antibodies -
Description
Rabbit monoclonal [EPR22994-255] to IL-8 -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: U-87 MG treated with 1µM Thapsigargin for 24 hours, whole cell lysate; Wild-type PC-3 LPS-treated (2 µg/ml, 6 h) with Brefeldin A (ab120299) (5 µg/ml, 5 h) cell lysate; His-tagged human recombiant IL-8 protein (aa21-99).
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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% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22994-255 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab235584 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 |
1/1000. Predicted molecular weight: 11 kDa.
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| Notes |
|---|
|
WB
1/1000. Predicted molecular weight: 11 kDa. |
Target
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Function
IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively. -
Sequence similarities
Belongs to the intercrine alpha (chemokine CxC) family. -
Post-translational
modificationsSeveral N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 3576 Human
- Omim: 146930 Human
- SwissProt: P10145 Human
- Unigene: 624 Human
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Alternative names
- (Ala-IL-8)77 antibody
- (Ser-IL-8)72 antibody
- 9E3 antibody
see all
Images
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Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)All lanes : Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution
Lane 1 : Wild-type PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
Lane 2 : Wild-type PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
Lane 3 : CXCL8 knockout PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
Lane 4 : CXCL8 knockout PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
Lane 5 : A431 cell lysate
Lane 6 : HCT116 cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 10 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab235584 observed at 10 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab235584 was shown to react with IL-8 in wild-type PC-3 cells in Western blot with loss of signal observed in CXCL8 knockout cell line ab273743 (knockout cell lysate ab275520). Wild-type PC-3 and CXCL8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab235584 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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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![Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)](https://www.abcam.com/ps/products/235/ab235584/Images/ab235584-407604-anti-il8-antibody-epr22994255-western-blot-il8.jpg)
Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution + His-tagged human recombiant IL-8 protein (aa21-99) 10 ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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![Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)](https://www.abcam.com/ps/products/235/ab235584/Images/ab235584-406521-anti-il8-antibody-epr22994255-western-blot-glioblastomaastrocytoma.jpg)
Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)All lanes : Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution
Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lane 2 : U-87 MG treated with 1µM Thapsigargin for 24 hours, whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes
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![Anti-IL-8 antibody [EPR22994-255] (ab235584)](https://www.abcam.com/ps/products/235/ab235584/Images/ab235584-4-benefits-of-recombinant-antibodies.png)
Anti-IL-8 antibody [EPR22994-255] (ab235584)
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 (2)
ab235584 has been referenced in 2 publications.
- Ye F et al. Osteogenic differentiation of mesenchymal stem cells promotes c-Jun-dependent secretion of interleukin 8 and mediates the migration and differentiation of CD4+ T cells. Stem Cell Res Ther 13:58 (2022). PubMed: 35123547
- Zhao H & He Y The Inhibitory Effect of Lysophosphatidylcholine on Proangiogenesis of Human CD34+ Cells Derived Endothelial Progenitor Cells. Front Mol Biosci 8:682367 (2021). PubMed: 34179086