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BV421 Rat Anti-Mouse CD103
BV421 Rat Anti-Mouse CD103

Multicolor flow cytometric analysis of CD103 expression on mouse lymph node cells (left and center panels). Lymph node cells from a C57BL/6 mouse were stained with Alexa Fluor® 488 Hamster Anti-Mouse CD3e antibody (Cat. No. 557666) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control  (Cat. No. 562602; Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse CD103 antibody (Cat. No. 562771; Center Panel). Two-color dot plots showing the coexpressed levels of CD103 (or Ig Isotype Control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Immunohistofluorescent analysis of CD103 expression by cells within C57BL/6 mouse intestine (right panel). A mouse intestine cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Rat Anti-Mouse CD103 antibody (Cat. No. 562771, pseudo-colored green). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored red). Images were captured on a standard four laser confocal microscope. Original magnification, 40x

Multicolor flow cytometric analysis of CD103 expression on mouse lymph node cells (left and center panels). Lymph node cells from a C57BL/6 mouse were stained with Alexa Fluor® 488 Hamster Anti-Mouse CD3e antibody (Cat. No. 557666) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control  (Cat. No. 562602; Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse CD103 antibody (Cat. No. 562771; Center Panel). Two-color dot plots showing the coexpressed levels of CD103 (or Ig Isotype Control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Immunohistofluorescent analysis of CD103 expression by cells within C57BL/6 mouse intestine (right panel). A mouse intestine cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Rat Anti-Mouse CD103 antibody (Cat. No. 562771, pseudo-colored green). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored red). Images were captured on a standard four laser confocal microscope. Original magnification, 40x

Product Details
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BD Horizon™
Itgae; Integrin alpha-E; αE; alpha-E1; ITAE; Integrin αIEL chain; aM290
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse Intestinal Epithelial Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
16407
AB_2737783
Aqueous buffered solution containing BSA and ≤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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. 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.
  8. 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.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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Antibody Details
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M290

The M290 antibody specificaly binds to CD103, the α chain of αIELβ7 integrin. CD103 has a unique and fairly restricted tissue distribution. It is expressed on almost all intestinal intraepithelial lymphocytes (IEL), dendritic epidermal T cells (DEC), subpopulations of peripheral T cells, and distinct subsets of fetal, neonatal, and adult thymocytes. E-cadherin is the epithelial cell ligand for αIELβ7 integrin. The ordered expression of αIEL during thymocyte development (which occurs under the influence of the thymic epithelium), high level of αIEL expression on peripheral T cells in epithelial tissues (IEL and DEC), and expression of CD103 on a subset of CD8+ lymphocytes responding to allogeneic epithelial cells, suggest that αIELβ7 integrin may have a common role in the interactions of T lymphocytes with epithelia during T-cell maturation and effector functions. CD103 is thought to play a role in allograft rejection. The M290 antibody is reported to efficiently inhibit αIELβ7-mediated adhesion in in vitro assays.

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.

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Format Details
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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.
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BV421
Violet 405 nm
407 nm
423 nm
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Citations & References
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Development References (7)

  1. Andrew DP, Rott LS, Kilshaw PJ, Butcher EC. Distribution of alpha 4 beta 7 and alpha E beta 7 integrins on thymocytes, intestinal epithelial lymphocytes and peripheral lymphocytes. Eur J Immunol. 1996; 26(4):897-905. (Clone-specific: Flow cytometry). View Reference
  2. Feng Y, Wang D, Yuan R, Parker CM, Farber DL, Hadley GA. CD103 expression is required for destruction of pancreatic islet allografts by CD8(+) T cells. J Exp Med. 2002; 196(7):877-886. (Biology). View Reference
  3. Hadley GA, Bartlett ST, Via CS, Rostapshova EA, Moainie S. The epithelial cell-specific integrin, CD103 (alpha E integrin), defines a novel subset of alloreactive CD8+ CTL. J Immunol. 1997; 159(8):748-3756. (Biology). View Reference
  4. Karecla PI, Bowden SJ, Green SJ, Kilshaw PJ. Recognition of E-cadherin on epithelial cells by the mucosal T cell integrin alpha M290 beta 7 (alpha E beta 7). Eur J Immunol. 1995; 25(3):852-856. (Clone-specific: Blocking). View Reference
  5. Kilshaw PJ, Baker KC. A unique surface antigen on intraepithelial lymphocytes in the mouse. Immunol Lett. 1988; 18(2):149-154. (Immunogen: Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
  6. Kilshaw PJ, Murant SJ. A new surface antigen on intraepithelial lymphocytes in the intestine. Eur J Immunol. 1990; 20(10):2201-2207. (Clone-specific: Immunoprecipitation). View Reference
  7. Kilshaw PJ, Murant SJ. Expression and regulation of beta 7(beta p) integrins on mouse lymphocytes: relevance to the mucosal immune system. Eur J Immunol. 1991; 21(10):2591-2597. (Clone-specific: Immunohistochemistry). View Reference
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