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Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin)
Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin)
Two-color flow cytometric analysis of CD300LG (Nepmucin) expression on mouse peritoneal exudate cells. BALB/c mouse peritoneal exudate cells (PECs) were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution and were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557906; Left Plot) or Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin) antibody (Cat. No. 566997; Right Plot) at 0.5 µg/test. Bivariate pseudocolor density plots showing the correlated expression of CD300LG (Nepmucin) [or Ig Isotype control staining] versus F4/80 were derived from gated events with the forward and side light-scatter characteristics of total viable PECs. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD300LG (Nepmucin) expression on mouse peritoneal exudate cells. BALB/c mouse peritoneal exudate cells (PECs) were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution and were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557906; Left Plot) or Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin) antibody (Cat. No. 566997; Right Plot) at 0.5 µg/test. Bivariate pseudocolor density plots showing the correlated expression of CD300LG (Nepmucin) [or Ig Isotype control staining] versus F4/80 were derived from gated events with the forward and side light-scatter characteristics of total viable PECs. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
CLM-9; CMRF-35-like molecule-9; Cd300lg; Clm9
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse Nepmucin extracellular domain
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869997
Aqueous buffered solution containing ≤0.09% sodium azide.
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566997 Rev. 1
Antibody Details
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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.

566997 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566997 Rev.1
Citations & References
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View product citations for antibody "566997" on CiteAb

Development References (4)

  1. 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
  2. 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
  3. 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
  4. 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
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566997 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

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.