Description
The 1G8 monoclonal antibody specifically recognizes Endothelial cell-selective adhesion molecule (ESAM). ESAM is a ~55 kDa single-pass type I transmembrane glycoprotein that is encoded by ESAM (Endothelial cell-specific adhesion molecule) which belongs to the immunoglobulin supergene superfamily (IgSF). ESAM contains an IgV-type and IgC2-type domain in its extracellular region followed by a transmembrane sequence and cytoplasmic tail. ESAM is strongly expressed on platelets, megakaryocytes, and endothelial cells at interendothelial cell junctions as well as dendritic cells. Through homophilic interactions, this interendothelial cell adhesion molecule may play roles in regulating vascular permeability and in the extravasation of leucocytes, eg, neutrophils, through blood vessel walls. ESAM is also reportedly expressed on hematopoietic stem cells (HSCs). ESAM-positive cell populations are enriched for multipotent myeloid erythroid progenitors and primitive progenitors with lymphopoietic activity.
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
6. An isotype control should be used at the same concentration as the antibody of interest.
7. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Specifications

General

BrandBD OptiBuild™
Alternative Name1G8 Antigen; Esam; Esam1; endothelial cell-selective adhesion molecule
ReactivityMouse (Tested in Development)
IsotypeRat LEW, also known as Lewis IgG2a, κ
ImmunogenMouse endothelial Cell Line
ApplicationFlow cytometry (Qualified)
Concentration0.2 mg/ml
RRIDAB_3687851
Storage BufferAqueous buffered solution containing ≤0.09% sodium azide.
Regulatory StatusRUO