Human ITGA2 (CD49b) knockout HeLa cell lysate (ab257262)
Overview
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Product name
Human ITGA2 (CD49b) knockout HeLa cell lysate
See all CD49b kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 4. -
Passage number
Knockout validation
Sanger Sequencing, 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.
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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260092 - Human ITGA2 knockout HeLa cell lysate 1 x 100µg ab255552 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
Target
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Relevance
Human CD49b is an integrin alpha 2 subunit that forms a heterodimer with beta 1 integrin and functions as an adhesion molecule. CD49b is a 160 kDa glycoprotein that noncovalently associates with the 130 kDa integrin beta 1 subunit (CD29) to form glycoprotein Ia/IIa (when expressed on platelets), and very late (activation) antigen 2 ('VLA2') when found on T cells. CD49b is expressed by platelets, approximately 50% of monocytes, long term cultivated T cells and most adherent cell lines. Integrin alpha 2/beta 1 is a receptor for laminin, collagen, collagen C propeptides, fibronectin and E cadherin. It recognizes the proline hydroxylated sequence G-F-P-G-E-R in collagen. It is responsible for adhesion of platelets and other cells to collagens, modulation of collagen and collagenase gene expression, force generation and organization of newly synthesized extracellular matrix. -
Cellular localization
Cell Membrane; Single pass type I membrane protein. -
Alternative names
- BR
- CD49 antigen like family member B
- CD49B
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260092 - Human ITGA2 knockout HeLa cell lysate 1 x 100µg ab255552 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
Images
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Western blot - Human ITGA2 knockout HeLa cell lysate (ab257262)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ITGA2 knockout HeLa cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab181549 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.
ab181549 Anti-Integrin alpha 2 antibody [EPR17349] was shown to specifically react with Integrin alpha 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264819 (knockout cell lysate ab257262) was used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab181549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.
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Western blot - Human ITGA2 knockout HeLa cell lysate (ab257262)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ITGA2 knockout HeLa cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab109432 observed at 129 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109432 Anti-Integrin alpha 2 antibody [EPR5789] was shown to specifically react with Integrin alpha 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264819 (knockout cell lysate ab257262) was used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab109432 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.
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Sanger Sequencing - Human ITGA2 knockout HeLa cell lysate (ab257262)Homozygous: 14 bp deletion in exon 4
