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Alexa Fluor™ 488 Rat Anti-Mouse CD199 (CCR9)

BD Pharmingen™ Alexa Fluor™ 488 Rat Anti-Mouse CD199 (CCR9)

Clone 9B1 (also known as 9B-1)

(RUO)
Alexa Fluor™ 488 Rat Anti-Mouse CD199 (CCR9)
Two-color flow cytometric analysis of CD199 (CCR9) expression on Mouse thymocytes. BALB/c Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat No. 553035/561093) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse CD199 (CCR9) antibody (Cat. No. 568790/568791; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD199 (CCR9) [or Ig Isotype control staining] versus CD8a was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD199 (CCR9) expression on Mouse thymocytes. BALB/c Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat No. 553035/561093) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse CD199 (CCR9) antibody (Cat. No. 568790/568791; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD199 (CCR9) [or Ig Isotype control staining] versus CD8a was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
C-C chemokine receptor type 9; CC-CKR-9; CCR-9; Ccr9; GPR-9-6
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Mouse CCR9 N-terminal Peptide
Flow cytometry (Routinely Tested)
0.2 mg/ml
12769
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 the dye under optimum conditions and unreacted dye 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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
Antibody Details
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9B1

The 9B1 monoclonal antibody specifically binds to CD199 which is also known as Chemokine (C-C motif) receptor 9 (CCR9) or C-C chemokine receptor type 9 (C-C CKR-9). CD199 is a ~42 kDa seven-transmembrane protein and member of the G protein-coupled receptor 1 family. CD199 binds to CCL25, which is likewise known as thymus-expressed chemokine (TECK) or small inducible chemokine 25 (Scya25). CCL25 is selectively and constitutively expressed in the thymic cortex and small intestinal epithelium. CD199 (CCR9) is highly expressed on CD4+CD8+ double-positive thymocytes and mature naïve CD8+ T cells, but not naïve CD4+ T cells, in the peripheral lymphoid organs. CD199 (CCR9) is also expressed on small intestinal B cells, and on subsets of memory CD4+ T cells and CD8+ T cells, and dendritic cells. The CCR9/CCL25 pathway plays important roles in T cell development and gut-associated immune functions. It is especially involved in the recruitment of CD8αα+ intraepithelial lymphocytes (IELs) and the homing of other lymphocytes to the gut. Dysregulation of this pathway is associated with inflammatory responses, such as inflammatory bowel disease (IBD) and celiac disease.

Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
Citations & References
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View product citations for antibody "568791" on CiteAb

Development References (9)

  1. Drakes ML, Stiff PJ, Blanchard TG. Inverse relationship between dendritic cell CCR9 expression and maturation state. Immunology. 2009; 127(4):466-476. (Biology). View Reference
  2. Feng N, Jaimes MC, Lazarus NH, et al. Redundant role of chemokines CCL25/TECK and CCL28/MEC in IgA+ plasmablast recruitment to the intestinal lamina propria after rotavirus infection. J Immunol. 2006; 176(10):5749-5759. (Biology). View Reference
  3. Hsu FC, Shapiro MJ, Dash B, et al. An Essential Role for the Transcription Factor Runx1 in T Cell Maturation.. Sci Rep. 2016; 6:23533. (Clone-specific: Flow cytometry). View Reference
  4. McGuire HM, Vogelzang A, Ma CS, et al. A subset of interleukin-21+ chemokine receptor CCR9+ T helper cells target accessory organs of the digestive system in autoimmunity. Immunity. 2011; 34(4):602-615. (Biology). View Reference
  5. Miething C, Scuoppo C, Bosbach B, et al. PTEN action in leukaemia dictated by the tissue microenvironment.. Nature. 2014; 510(7505):402-6. (Clone-specific: Flow cytometry). View Reference
  6. Mirlekar B, Ghorai S, Khetmalas M, Bopanna R, Chattopadhyay S. Nuclear matrix protein SMAR1 control regulatory T-cell fate during inflammatory bowel disease (IBD).. Mucosal Immunol. 2015; 8(6):1184-200. (Clone-specific: Flow cytometry). View Reference
  7. Rivera-Nieves J, Ho J, Bamias G, et al. Antibody blockade of CCL25/CCR9 ameliorates early but not late chronic murine ileitis. Gastroenterology. 2006; 131(5):1518-1529. (Immunogen: Functional assay). View Reference
  8. Wendland M, Czeloth N, Mach N, et al. CCR9 is a homing receptor for plasmacytoid dendritic cells to the small intestine. Proc Natl Acad Sci U S A. 2007; 104(15):6347-6352. (Biology). View Reference
  9. Zaballos A, Gutierrez J, Varona R, Ardavin C, Marquez G. Cutting edge: identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. J Immunol. 1999; 162(10):5671-5675. (Biology). View Reference
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