Multicolor flow cytometric analysis of LPAM-1 expression on C57BL/6 mouse bone marrow cells. Bone marrow cells were stained with Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557683) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse LPAM-1 antibody (Cat. No. 562668; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of LPAM-1 (or Ig isotype control staining) versus CD45R/B220 for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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Multicolor flow cytometric analysis of LPAM-1 expression on C57BL/6 mouse bone marrow cells. Bone marrow cells were stained with Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557683) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse LPAM-1 antibody (Cat. No. 562668; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of LPAM-1 (or Ig isotype control staining) versus CD45R/B220 for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
Itga4Itgb7; α4β7 Integrin; alpha 4 beta 7 integrin; CD49dβ7
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2a, κ
AKR/Cum mouse T lymphoma line TK1
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2737709
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™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- CF™ is a trademark of Biotium, Inc.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
PE-CF594 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562302
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Alexa Fluor® 647 Rat Anti-Mouse CD45R RUO
Size
0.1 mg
Cat No.
557683
562668 Rev. 1
Antibody Details
DATK32
The DATK32 monoclonal antibody specifically binds to an epitope specific for the α4β7 integrin heterodimer. α4β7 (LPAM-1) is expressed on most mature lymphocytes and on small subsets of thymic and bone marrow cells. It interacts with several ligands, including VCAM-1 (CD106), fibronectin, and MAdCAM-1. DATK32 antibody induces α4β7-dependent lymphocyte aggregation, but it inhibits other α4β7- mediated lymphocyte adhesion events, including binding to fibronectin, MAdCAM-1, and VCAM-1 (CD106).
This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
562668 Rev. 1
Format Details
PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
562668 Rev.1
Citations & References
Development References (5)
-
Andrew DP, Berlin C, Honda S, et al. Distinct but overlapping epitopes are involved in alpha 4 beta 7-mediated adhesion to vascular cell adhesion molecule-1, mucosal addressin-1, fibronectin, and lymphocyte aggregation. J Immunol. 1994; 153(9):3847-3861. (Immunogen: Blocking, Induction). View Reference
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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: Blocking). View Reference
-
Berlin C, Berg EL, Briskin MJ, et al. Alpha 4 beta 7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell. 1993; 74(1):185-195. (Clone-specific: Blocking). View Reference
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Hamann A, Andrew DP, Jablonski-Westrich D, Holzmann B, Butcher EC. Role of alpha 4-integrins in lymphocyte homing to mucosal tissues in vivo. J Immunol. 1994; 152(7):3282-3293. (Clone-specific). View Reference
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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. (Biology). View Reference
562668 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.
