ICAM1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Alternative names
Antigen identified by monoclonal antibody BB2, BB 2, CD 54, CD_antigen=CD54, Cell surface glycoprotein P3.58, Human rhinovirus receptor, ICAM1_HUMAN, Intercellular adhesion molecule 1, MALA 2, Major group rhinovirus receptor, MyD 10, P3.58, Surface antigen of activated B cells, Surface antigen of activated B cells, BB2, intercellular adhesion molecule 1 (CD54), intercellular adhesion molecule 1 (CD54), human rhinovirus receptor
What's included?
Recommended products
ICAM1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ICAM1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
6 product images
Western blot - Human ICAM1 knockout HeLa cell lysate (ab256947)
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: ICAM1 knockout HeLa cell lysate 20 μg
Lane 3: A549 cell lysate 20 μg
Lane 4: Ramos cell lysate 20 μg
False colour image of Western blot: Anti-ICAM1 antibody [EP1442Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-ICAM1 antibody [EP1442Y] ab53013 was shown to bind specifically to ICAM1. A band was observed at 90 kDa (not observed by this antibody) in wild-type HeLa cell lysates with no signal observed at this size in Icam1 knockout cell line Human ICAM1 knockout HeLa cell line ab261742 (knockout cell lysate ab256947). The band observed in the knockout lysate lane below 90 kDa is likely to represent a truncated form of ICAM1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Icam1 knockout HeLa 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.All lanes: Western blot - Anti-ICAM1 antibody [EP1442Y] (Anti-ICAM1 antibody [EP1442Y] ab53013) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ICAM1 knockout HeLa cell lysate at 20 µg
Lane 3: A549 cell lysate at 20 µg
Lane 4: Ramos cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 90 kDa
Western blot - Human ICAM1 knockout HeLa cell lysate (ab256947)
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: ICAM1 knockout HeLa cell lysate 20 μg
Lane 3: A549 cell lysate 20 μg
Lane 4: Ramos cell lysate 20 μg
False colour image of Western blot: Anti-ICAM1 antibody [EPR4776] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-ICAM1 antibody [EPR4776] ab109361 was shown to bind specifically to ICAM1. A band was observed at 90 kDa in wild-type HeLa cell lysates with no signal observed at this size in Icam1 knockout cell line Human ICAM1 knockout HeLa cell line ab261742 (knockout cell lysate ab256947). The band observed in the knockout lysate lane below 90 kDa is likely to represent a truncated form of ICAM1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Icam1 knockout HeLa 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.All lanes: Western blot - Anti-ICAM1 antibody [EPR4776] (Anti-ICAM1 antibody [EPR4776] ab109361) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ICAM1 knockout HeLa cell lysate at 20 µg
Lane 3: A549 cell lysate at 20 µg
Lane 4: Ramos cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 90 kDa
Western blot - Human ICAM1 knockout HeLa cell lysate (ab256947)
Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lane 1: Wild-type HeLa cell lysate
Lane 2: ICAM1 knockout HeLa cell lysate
Lane 3: Ramos cell lysateLanes 1-3: Merged signal (red and green). Green - Anti-ICAM1 antibody [EPR24639-3] ab282575 observed at 90kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-ICAM1 antibody [EPR24639-3] ab282575 Anti-ICAM1 antibody [EPR24639-3] was shown to react with ICAM1 in wild-type Hela cells in Western blot. Loss of signal was observed when knockout cell line Human ICAM1 knockout HeLa cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE.
Anti-ICAM1 antibody [EPR24639-3] ab282575 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at 4oC overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Lane 1: Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2: ICAM1 knockout HeLa whole cell lysate
Lane 3: Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
SecondaryAll lanes: Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution
Western blot - Human ICAM1 knockout HeLa cell lysate (ab256947)
Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:ICAM1 knockout HeLa cell lysate (20 ug)
Lane 3:Raji cell lysate (20 ug)
Lane 4:Ramos cell lysate (20 ug)Anti-ICAM1 antibody [EP1442Y] ab53013 was shown to specifically react with ICAM1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human ICAM1 knockout HeLa cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE. Anti-ICAM1 antibody [EP1442Y] ab53013 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Sanger Sequencing - Human ICAM1 knockout HeLa cell lysate (ab256947)
Allele-2: 1 bp insertion in exon 2
Sanger Sequencing - Human ICAM1 knockout HeLa cell lysate (ab256947)
Allele-1: Insertion of the selection cassette in exon 2
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