Human IL-6 knockout A549 cell lysate
Be the first to review this product! Submit a review
|
(0 Publication)
IL6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 3.
View Alternative Names
B cell differentiation factor, B-cell stimulatory factor 2, BSF-2, CTL differentiation factor, Cytotoxic T cell differentiation factor, HSF, Hepatocyte stimulating factor, Hepatocyte stimulatory factor, Hybridoma growth factor, Hybridoma growth factor Interferon beta-2, Hybridoma plasmacytoma growth factor, IFN-beta-2, IFNB2, IL6_HUMAN, Interferon beta-2, Interleukin 6 (interferon beta 2), Interleukin BSF 2, Interleukin-6
- WB
Lab
Western blot - Human IL-6 knockout A549 cell lysate (AB275501)
Lane 1 : Wild-type A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
Lane 2 : Wild-type A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
Lane 3 : IL-6 knockout A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
Lane 4 : IL-6 knockout A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
False colour image of Western blot : Anti-IL-6 antibody [EPR22565-204] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab233551 was shown to bind specifically to IL-6. A band was observed at 25 kDa in wild-type A549 cell lysates with no signal observed at this size in IL6 knockout cell line ab273751 (knockout cell lysate ab275501). To generate this image, wild-type and IL6 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-IL-6 antibody [EPR22565-204] (<a href='/en-us/products/primary-antibodies/il-6-antibody-epr22565-204-ab233551'>ab233551</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 ug/mL, final 6 h) cell lysate at 30 µg
Lane 2:
Wild-type A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 ug/mL, final 6 h) cell lysate at 30 µg
Lane 3:
IL-6 knockout A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 ug/mL, final 6 h) cell lysate at 30 µg
Lane 4:
IL-6 knockout A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 ug/mL, final 6 h) cell lysate at 30 µg
Predicted band size: 23 kDa
Observed band size: 25 kDa
false
- WB
Lab
Western blot - Human IL-6 knockout A549 cell lysate (AB275501)
Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 2 : Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 4 : IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab214429 observed at 25 kDa, possible non-specific binding at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab214429 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type A549 and IL-6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214429 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IL-6 antibody [EPR20653] (<a href='/en-us/products/primary-antibodies/il-6-antibody-epr20653-ab214429'>ab214429</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 2:
Wild-type A549 IL-1β (<a href='/en-us/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Lane 2:
Western blot - Human IL-6 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-il-6-knockout-a549-cell-line-ab273751'>ab273751</a>)
Lane 3:
IL-6 knockout A549 Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 4:
IL-6 knockout A549 IL-1β (<a href='/en-us/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Predicted band size: 23 kDa
Observed band size: 25 kDa,35 kDa
false
- WB
Lab
Western blot - Human IL-6 knockout A549 cell lysate (AB275501)
Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 2 : Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 4 : IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab233706 observed at 25 kDa, possible non-specific binding at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab233706 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab233706 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IL-6 antibody [EPR21711] (<a href='/en-us/products/primary-antibodies/il-6-antibody-epr21711-ab233706'>ab233706</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 2:
Wild-type A549 IL-1β (<a href='/en-us/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Lane 2:
Western blot - Human IL-6 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-il-6-knockout-a549-cell-line-ab273751'>ab273751</a>)
Lane 3:
IL-6 knockout A549 Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 4:
IL-6 knockout A549 IL-1β (<a href='/en-us/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Predicted band size: 23 kDa
Observed band size: 25 kDa,35 kDa,40 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human IL-6 knockout A549 cell lysate (AB275501)
Allele-1 : 97 bp deletion in exon 3.
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
Treatments:
Human IL-6 knockout A549 cell lysate - Brefeldin A (5 μg/ml, 6h)
Human wild-type A549 cell lysate - Brefeldin A (5 μg/ml, 6h)
Human IL-6 knockout A549 cell lysate - IL-1ß (20 ng/ml, 24h) and Brefeldin A (5 μg/ml, during the last 6h of IL-1ß treatment)
Human wild-type A549 cell lysate - IL-1ß (20 ng/ml, 24h) and Brefeldin A (5 μg/ml, during the last 6h of IL-1ß treatment)
REACH authorisation
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.
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. 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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
IL-6 influences immune regulation and acts as part of the acute phase response. It stimulates the production of acute-phase proteins and supports the differentiation of B cells into antibody-producing cells. IL-6 is not known to be part of a larger complex acting primarily as a single entity in signal transduction. Moreover IL-6 impacts the metabolism of iron and bone homeostasis showing its multifunctional nature.
Pathways
IL-6 forms an integral part of several signaling routes particularly the JAK-STAT pathway. In this context IL-6 interacts with signal transducer proteins like STAT3 to transmit signals from the cell surface to the nucleus affecting gene expression. Another important pathway is the MAPK pathway through which IL-6 influences cell proliferation and survival. These interactions reflect IL-6's diverse effects in cellular processes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com