Integrins are heterodimers composed of noncovalently associated transmembrane a and b subunits. The 16 a and 8 b subunits heterodimerize to produce more than 20 different receptors. Most integrin receptors bind ligands that are components of the extracellular matrix, including Fibronectin, collagen and vitronectin. Certain integrins can also bind to soluble ligands, such as fibrinogen, or to counterreceptors on adjacent cells such as the intracellular adhesion molecules (ICAMs), leading to aggregation of cells. Ligands serve to cross-link or cluster integrins by binding to adjacent integrin receptors; both receptor clustering and ligand occupancy are necessary for the activation of integrin-mediated responses. In addition to mediating cell adhesion and cytoskeletal organization, integrins function as signaling receptors. Signals transduced by integrins play a role in many biological processes, including cell growth, differentiation, migration and apoptosis.
Background References
1. Coughlan L et al. Combined fiber modifications both to target a(v) (6) and detarget the coxsackievirus-adenovirus receptor improve virus toxicity profiles in vivo but fail to improve antitumoral efficacy relative to adenovirus serotype 5. Hum Gene Ther 23:960-79 (2012).
2. Marsh D et al. alpha vbeta 6 Integrin promotes the invasion of morphoeic basal cell carcinoma through stromal modulation. Cancer Res 68:3295-303 (2008).
Western blot analysis of Integrin alpha V on different lysates with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/2,000 dilution.
Lane 1: A549 cell lysate (10 µg/Lane) Lane 2: HeLa cell lysate (10 µg/Lane) Lane 3: NIH/3T3 cell lysate (10 µg/Lane) Lane 4: C6 cell lysate (10 µg/Lane) Lane 5: Rat heart tissue lysate (20 µg/Lane)
Predicted band size: 116 kDa Observed band size: 125/135 kDa
Exposure time: 25 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-15) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of Integrin alpha V with anti-Integrin alpha V antibody (ET1610-15) at 1/2,000 dilution.
Lane 1: Wild-type A549 whole cell lysate. Lane 2/3: Integrin alpha V knockdown A549 whole cell lysate.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-Integrin alpha V antibody (ET1610-15, 1/2,000) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Integrin alpha V on different lysates with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/5,000 dilution.
Lane 1: A549 cell lysate Lane 2: HeLa cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 116 kDa Observed band size: 130 kDa
Exposure time: 4 minutes;
6% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-15) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Flow cytometric analysis of Integrin alpha V was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-15, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-15) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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