BV421 Rat Anti-Mouse CD300LG (Nepmucin)

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BD OptiBuild™ BV421 Rat Anti-Mouse CD300LG (Nepmucin)

Name: BV421 Rat Anti-Mouse CD300LG (Nepmucin)
Brand: BD OptiBuild™
SKU: 752372

Clone ZAQ2

(RUO)

Two-color flow cytometric analysis of CD300LG (Nepmucin) expression on Mouse peritoneal exudate cells. Mouse peritoneal exudate cells (PEC) from BALB/c mice were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution. PECs were incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. Cells were then stained with PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and with either BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left panel) or BD OptiBuild™ BV421 Rat Anti-Mouse CD300LG (Nepmucin) antibody (Cat. No. 752372; Right Panel) at 0.125ug/test. The two-color flow cytometric contour plot showing the correlated expression of CD300LG (Nepmucin) [or Ig Isotype control staining] versus F4/80 was derived from gated events with the forward and side light-scatter characteristics of total viable PECs. Flow cytometry and data analysis were performed using a BD FACSCelesta™ 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: CLM-9; CMRF-35-like molecule-9; Cd300lg; Clm9

Reactivity: Mouse (Tested in Development)

Isotype: Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ

Immunogen: Mouse Nepmucin extracellular domain

Application: Flow cytometry (Qualified)

Concentration: 0.2 mg/ml

RRID: AB_2875889

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

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Lysing Buffer RUO

Size 100 mL Cat No. 555899

View Details

752372 Rev. 2

Antibody Details

ZAQ2

The ZAQ2 monoclonal antibody specifically recognizes CD300LG, a microvascular endothelial adhesion molecule that belongs to the CD300 antigen like family. CD300LG is also known as Nepmucin (a not expressed in Peyer's patches mucin) and CMRF35-like molecule 9 (CLM-9). Four isoforms of this type I transmembrane glycoprotein have been described that are encoded by Cd300lg. The extracellular region of this sialomucin contains one IgV-like domain and one mucin-like domain followed by a transmembrane region and an intracellular tail. The ZAQ2 antibody recognizes the Ig domain of CD300LG (Nepmucin). This sialomucin is variably expressed on the luminal surfaces of endothelial cells lining small arterioles, venules and capillaries in many tissues. Although CD300LG (Nepmucin) protein is expressed by high endothelial venules (HEV) in lymph nodes, it is not expressed by Peyer's patches HEV. Nepmucin mRNA expression has also been observed for some myeloid lineage cell lines as well as CD11c+ dendritic cells, but not purified T and B lymphocytes. HEV-associated CD300LG (Nepmucin) mediates homotypic interactions between endothelial cells. This sialomucin also supports L-selectin-dependent lymphocyte rolling and heterophilic binding of lymphocytes by endothelial cells leading to transendothelial migration (TEM) of lymphocytes. The ZAQ2 antibody can reportedly inhibit lymphocyte TEM. CD300LG (Nepmucin) expression on microvascular endothelial cells is affected by local factors generated in tissues, including sites undergoing inflammation or tumor cell infiltration. This may in turn affect the levels of lymphocytic infiltration influenced by this sialomucin.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

752372 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

752372 Rev.2

Citations & References

Development References (4)

Jin S, Umemoto E, Tanaka T, et al. Nepmucin/CLM-9, an Ig domain-containing sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.. FEBS Lett. 2008; 582(20):3018-24. (Clone-specific: Functional assay, Immunofluorescence, Inhibition). View Reference
Umemoto E, Hayasaka H, Bai Z, et al. Novel regulators of lymphocyte trafficking across high endothelial venules.. Crit Rev Immunol. 2011; 31(2):147-69. (Biology). View Reference
Umemoto E, Takeda A, Jin S, et al. Dynamic changes in endothelial cell adhesion molecule nepmucin/CD300LG expression under physiological and pathological conditions.. PLoS ONE. 2013; 8(12):e83681. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
Umemoto E, Tanaka T, Kanda H, et al. Nepmucin, a novel HEV sialomucin, mediates L-selectin-dependent lymphocyte rolling and promotes lymphocyte adhesion under flow.. J Exp Med. 2006; 203(6):1603-14. (Immunogen: ELISA, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). View Reference

View All (4)

752372 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.