Human CXCL8 knockout PC-3 cell lysate (ab275520)
Overview
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Product name
Human CXCL8 knockout PC-3 cell lysate
See all IL-8 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
Treatments:
Human CXCL8 knockout PC3 cell lysate - Brefeldin A (5 µg/ml, 5h)
Human wild-type PC3 cell lysate - Brefeldin A (5 µg/ml, 5h)
Human CXCL8 knockout PC3 cell lysate - LPS (2 µg/ml, 6h) and Brefeldin A (5 µg/ml, 5h)
Human wild-type PC3 cell lysate - LPS (2 µg/ml, 6h) and Brefeldin A (5 µg/ml, 5h) -
Parental Cell Line
PC3 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 22% 11 bp deletion, 24% 7 bp deletion, 54% 2 bp deletion in exon 2 -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277344 - Human CXCL8 knockout PC3 cell lysate - Brefeldin A treated 1 x 100µg ab277343 - Human CXCL8 knockout PC3 cell lysate - LPS + Brefeldin A treated 1 x 100µg ab277346 - Human wild-type PC3 cell lysate - Brefeldin A treated 1 x 100µg ab277345 - Human wild-type PC3 cell lysate - LPS + Brefeldin A treated 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Male
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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Alternative names
- (Ala-IL-8)77
- (Ser-IL-8)72
- 9E3
see all
Associated products
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Biochemicals
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275520 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 |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Western blot - Human CXCL8 knockout PC3 cell lysate (ab275520)
Lane 1: Wild-type PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate 30 μg
Lane 2: Wild-type PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate 30 μg
Lane 3: CXCL8 knockout PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate 30 μg
Lane 4: CXCL8 knockout PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate 30 μg
Lane 5: A431 cell lysate 30 μg
Lane 6: HCT116 cell lysate 30 μg
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 37kDa.
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. -
Next Generation Sequencing - Human CXCL8 knockout PC3 cell lysateAllele-1: 11bp deletion in exon 2.
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Next Generation Sequencing - Human CXCL8 knockout PC3 cell lysateAllele-2: 7bp deletion in exon 2.
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Next Generation Sequencing - Human CXCL8 knockout PC3 cell lysateAllele-3: 2bp deletion in exon 2.
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
References (0)
ab275520 has not yet been referenced specifically in any publications.