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Anti-ITGB2 Antibody EP1288Y
Also for ITGB2 (NM_000211)
|A synthetic phospho-peptide corresponding to residues surrounding Ser745 of human LFA-?2 was used as immunogen. The antibody only detects LFA-?2 phosphorylated on Serine 745.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:500; IP: 1:100; IHC-P: Use at an assay dependent concentration; ICC/IF: 1:250 - 1:500; FC: 1:1000
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens integrin, beta 2 (complement component 3 receptor 3 and 4 subunit) (ITGB2), transcript variant 1|
|CD18; LAD; LCAMB; LFA-1; MAC-1; MF17; MFI7|
|In the immune system, integrins have essential roles in leukocyte trafficking and function. These include immune cell attachment to endothelial and antigen-presenting cells, cytotoxicity, and extravasation into tissues (1). Integrin adhesion receptors transduce signals that control complex cell functions which require the regulation of gene expression, such as proliferation, differentiation and survival. Their intracellular domain has no catalytic function, indicating that interaction with other transducing molecules is crucial for integrin-mediated signaling. JAB1 (Jun activation domain-binding protein 1), a coactivator of the c-Jun transcription factor, has been identified as a protein that interacts with the cytoplasmic domain of the beta2 subunit of the alphaL/beta2 integrin LFA-1 (2). Patients with leukocyte adhesion molecule (CD11/CD18, beta 2 integrins) deficiency have structural defects in the common beta subunit (CD18), which prevent heterodimer formation and normal cell surface expression of these receptors, leading to life-threatening bacterial infections (3) |
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Western blot - CD18 (phospho S745) antibody [EP1288Y]; All lanes : Anti-CD18 (phospho S745) antibody [EP1288Y] at 1/500 dilution.Lane 1 : Jurkat cell lysate, untreated.Lane 2 : Jurkat cell lysate, treated with AP.Lysates/proteins at 10 µg per lane.Secondary.Goat anti-rabbit HRP labeled at 1/2000 dilution.Predicted band size : 85 kDa.Observed band size : 85 kDa.
Immunohistochemistry (Paraffin-embedded sections) - CD18 (phospho S745) antibody [EP1288Y]; Ab52920 (1:250) staining human CD18 in human tonsil tissue by immunohistochemistry using paraffin embedded tissue.
Immunocytochemistry/ Immunofluorescence - Anti-CD18 (phospho S745) antibody [EP1288Y]; ICC/IF image of ab52920 stained Raw 246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab52920 at 1/1000 dilution overnight at +4°C. The secondary antibody (green) was DyLight? 488 goat anti- rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Flow Cytometry - Anti-CD18 (phospho S745) antibody [EP1288Y]; Human peripheral blood lymphocytes stained with ab52920 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody for 30 min at 4°C. The secondary antibody used was Alexa Fluor? 488 goat anti-rabbit IgG (H&L) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1??/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy ??peripheral blood lymphocytes.