BV421 Rat Anti-Mouse ESAM

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BD OptiBuild™ BV421 Rat Anti-Mouse ESAM

Name: BV421 Rat Anti-Mouse ESAM
Brand: BV421 Rat Anti-Mouse ESAM
SKU: 752436

Clone 1G8 (RUO)

Flow cytometric analysis of ESAM expression on bEnd.3 cells. Cells from the Mouse bEnd.3 (Brain Endothelioma, ATCC® CRL-2299™) cell line were stained with BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse ESAM antibody (Cat. No. 752436; solid line histogram) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing ESAM expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD Celesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD OptiBuild™

Alternative Name: 1G8 Antigen; Esam; Esam1; endothelial cell-selective adhesion molecule

Reactivity: Mouse (Tested in Development)

Isotype: Rat LEW, also known as Lewis IgG2a, κ

Immunogen: Mouse endothelial Cell Line

Application: Flow cytometry (Qualified)

Concentration: 0.2 mg/ml

Entrez Gene ID: 69524

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

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.

Recommended Assay Procedures

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.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

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.
Researchers should determine the optimal concentration of this reagent for their individual applications.
An isotype control should be used at the same concentration as the antibody of interest.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

Companion Products

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.1 mg Cat No. 553141

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Lysing Buffer RUO

Size 100 mL Cat No. 555899

View Details

752436 Rev. 2

Antibody Details

1G8

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.

752436 Rev. 2

Format Details

BV421

The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV421

Excitation Source: Violet 405 nm

Excitation Max: 407 nm

Emission Max: 423 nm

752436 Rev.2

Citations & References

View product citations for antibody "752436" on CiteAb

Development References (7)

Duong CN, Nottebaum AF, Butz S, et al. Interference With ESAM (Endothelial Cell-Selective Adhesion Molecule) Plus Vascular Endothelial-Cadherin Causes Immediate Lethality and Lung-Specific Blood Coagulation.. Arterioscler Thromb Vasc Biol. 2020; 40(2):378-393. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Western blot). View Reference
Fujita K, Chakarov S, Kobayashi T, et al. Cell-autonomous FLT3L shedding via ADAM10 mediates conventional dendritic cell development in mouse spleen.. Proc Natl Acad Sci U S A. 2019; 116(29):14714-14723. (Clone-specific: Flow cytometry). View Reference
Hirata Ki, Ishida T, Penta K, et al. Cloning of an immunoglobulin family adhesion molecule selectively expressed by endothelial cells.. J Biol Chem. 2001; 276(19):16223-31. (Biology). View Reference
Kirkling ME, Cytlak U, Lau CM, et al. Notch Signaling Facilitates In Vitro Generation of Cross-Presenting Classical Dendritic Cells.. Cell Rep. 2018; 23(12):3658-3672.e6. (Clone-specific: Flow cytometry). View Reference
Lewis KL, Caton ML, Bogunovic M, et al. Notch2 receptor signaling controls functional differentiation of dendritic cells in the spleen and intestine.. Immunity. 2011; 35(5):780-91. (Biology). View Reference
Nasdala I, Wolburg-Buchholz K, Wolburg H, et al. A transmembrane tight junction protein selectively expressed on endothelial cells and platelets.. J Biol Chem. 2002; 277(18):16294-303. (Immunogen: Electron microscopy, ELISA, Flow cytometry, Fluorescence microscopy, Immunoaffinity chromatography, Immunofluorescence, Immunohistochemistry, Western blot). View Reference
Sudo T, Yokota T, Oritani K, et al. The endothelial antigen ESAM monitors hematopoietic stem cell status between quiescence and self-renewal.. J Immunol. 2012; 189(1):200-10. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference

View All (7)

752436 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.