Specificity Not recommended for use with rat and mouse samples.
Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #ALR-001), and flow cytometry (labeled antibody).
Applications: FC
Direct flow cytometry
Live intact human promyelocytic leukemia (HL-60) cells (1:20-1:40).
Cell surface detection of LTB4R in live intact human promyelocytic leukemia HL-60 cell line:
Leukotrienes (LTs) are lipid mediators prominently exerting proinflammatory responses. They are divided in two classes: cysteinyl LTs (cysLTs) which have a thioester linkage and LTB41. LTB4, a chemoattractant, plays a prominent role in the recruitment and activation of leukocytes2.
LTB4 acts through two receptors which belong to the G-protein coupled receptor superfamily: BLT1 and BLT2. BLT1 displays high affinity for LTB4 and is highly expressed in leukocytes including granulocytes, macrophages, mast cells, dendritic cells and effector T cells3 and is expressed to much lower levels in the spleen, thymus, heart, skeletal muscle brain and liver2-4. BLT2 on the other hand is a low affinity receptor which does not display selectivity towards LTB45. Its expression is much more ubiquitous than that of BLT1. It is expressed in most human tissues, with the highest expression being in the spleen, liver, ovary, and leukocytes2,5-8. The two receptors could couple and activate different G-proteins, depending on the cell type and the cellular events activating the receptors2. However, activation of BLT1 usually correlates with increasing intracellular concentrations of Ca2+.
The LTB4/BLT1 system is involved in many allergic reactions as well as asthma induced as an allergic response, where it plays a significant role in recruiting neutrophils and effector T cells into lungs as part of an inflammatory response induced by allergens1.
Application key:
CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot