b'AntibodiesAntibodies against the receptor ITGB1Interactions of cells amongst each other and with the extracellularA HeLa WT control HeLa Cas9 control HeLa ITGB KOenvironment are mediated by different families of receptors. In addition to mediating cell adhesion and motility, these receptors also guide many cellular processes including growth and differentiation. A growing MA5-13658 MA1-81135body of evidence also suggests that these proteins help facilitate the development and progression of cancer, whereby alterations in their expression may bring about tumor malignancy, invasiveness, and metastasis. Integrins are a family of transmembrane receptors that have roles in cell cycle regulation, cytoskeleton organization, and recruitmentBof new receptors to the cell membrane. They work in conjunction with proteins such as cadherins, selectins, and syndecans to mediate cellcell and cellmatrix interactions.Integrin beta-1 (ITGB1 or CD29) belongs to a class of integrins that bind to collagen. These integrins are heterodimers and have large extracellular and short intracellular domains. ITGB1 interacts with various integrin alpha chains to form receptors for collagen, fibrinogen, fibronectin, osteopontin, VCAM1, and vitronectin among others. It is thought to play a role in endothelial motility and angiogenesis,Figure 3. Antibodies against ITGB1 tested for specificity using CRISPR-Cas9mediated knockout of ITGB1. Knockout of ITGB1 was achieved by CRISPR-Cas9 thereby inciting an interest in tumor progression and cancer genome editing using the InvitrogenLentiArrayLentiviral sgRNA (Cat. No. A32042, research. Additionally, ITGB1 is known to be involved in cell adhesion,Assay ID CRISPR626332_LV and CRISPR626355_LV) and the Invitrogen LentiArray invadopodia formation, and matrix degradation. Cas9 Lentivirus (Cat. No. A32064). Immunofluorescence (IF) analysis was performed on wild-type (WT) HeLa cells, Cas9-expressing HeLa cells, and HeLa ITGB1 knockout (KO) cells. (A) Cells were fixed, permeabilized, and labeled with the Invitrogen ITGB1 Invitrogen antibodies against ITGB1 are highly cited and tested forMonoclonal Antibody (4B7R) (Cat. No. MA5-13658, 1:100 dilution), followed by the applications such as WB, IF, IHC, and ELISA. Specificity verificationInvitrogen Goat AntiMouse IgG (H+L), Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Cat. No. A32723, 1:2,000, green). Nuclei (blue) were using techniques such as knockout via CRISPR-Cas9 genome editing stained using the InvitrogenProLongDiamond Antifade Mountant with DAPI (Cat. No. further ensures that the antibody binds to the intended target. Figure 3P36962), and Invitrogen Rhodamine Phalloidin (Cat. No. R415, 1:300) was used for showcases two such antibodies for ITGB1 (Cat. No. MA5-13658cytoskeletal F-actin (red) staining. Loss of signal upon CRISPR-Cas9mediated knockout confirms that the antibody is specific to ITGB1. (B) Cells were fixed, permeabilized, and and MA1-81135), verified using CRISPR-Cas9 genome editing forlabeled with the InvitrogenITGB1 Monoclonal Antibody (JB1B) (Cat. No. MA1-81135, immunofluorescence applications in HeLa cells. 1:100 dilution), followed by the Goat AntiMouse IgG (H+L), Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Cat. No. A32723, 1:2,000, green). Nuclei (blue) were stained using the ProLong Diamond Antifade Mountant with DAPI (Cat. No. P36962), and Rhodamine Phalloidin (Cat. No. R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal upon CRISPR-Cas9mediated knockout confirms that the antibody is specific to ITGB1. All images were captured at 60x magnification.Learn more about knockout verification at thermofisher.com/antibodyvalidation16 Inside the Cell//Issue No. 12Contents '