BD Pharmingen™ PE Rat Anti-Mouse Active Integrin β7
Clone H3/B7 (also known as H3) (RUO)
Two-color flow cytometric analysis of Active Integrin β7 expression on Mouse lymph node cells. C57BL/6 Mouse lymph node cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), then washed in Hanks' Balanced Salt Solution (HBSS), and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141]. The cells were then stained in HBSS (Top Plots) or HBSS + Mn2+ (Bottom Plots) with BD OptiBuild™ BV421 Integrin β7 chain antibody (Cat No. 743787), and with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930/554689; Left Plots) or PE Rat Anti-Mouse Active Integrin β7 antibody (Cat. No. 571202/571203; Right Plots) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to the cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Active Integrin β7 (or Ig Isotype control staining) versus Integrin β7 chain were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leukocytes. 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 Active Integrin β7 expression on Mouse lymph node cells. C57BL/6 Mouse lymph node cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), then washed in Hanks' Balanced Salt Solution (HBSS), and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141]. The cells were then stained in HBSS (Top Plots) or HBSS + Mn2+ (Bottom Plots) with BD OptiBuild™ BV421 Integrin β7 chain antibody (Cat No. 743787), and with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930/554689; Left Plots) or PE Rat Anti-Mouse Active Integrin β7 antibody (Cat. No. 571202/571203; Right Plots) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to the cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Active Integrin β7 (or Ig Isotype control staining) versus Integrin β7 chain were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leukocytes. 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.
Preparation And Storage
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
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- An isotype control should be used at the same concentration as the antibody of interest.
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Companion Products
The H3 monoclonal antibody specifically recognizes the activated conformational form of the Integrin β7 which is known as Active Integrin β7. Integrin β7 is a 130 kDa single pass type I transmembrane glycoprotein that is encoded by Itgb7 (integrin beta 7). Integrin β7 noncovalently associates with either Integrin α4 or αE chains to form Integrin α4/β7 or Integrin αE/β7 heterodimers. Integrin β7 is activated to an adhesive form within these complexes in response to various stimuli including Mn2+ ions, cytokines, or chemokines. Integrin α4β7, also known as CD49d/β7 or LPAM-1, is expressed on B and T lymphocytes, NK cells and on subsets of thymocytes and bone marrow cells. Activated Integrin α4β7 binds to Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) which acts as a key vascular addressin for intestinal homing. It also binds to CD106 (VCAM-1) expressed on dendritic cells and endothelial cells and the extracellular matrix protein fibronectin which are involved in the adhesion and migration of lymphocytes and some hematopoietic progenitor cells. Integrin αE/β7 (also known as, CD103/β7 or αIEL/β7) is expressed on intestinal intraepithelial lymphocytes (IEL), dendritic epidermal T cells, T regulatory cells, and a subset of CD8+ T cells in lymph nodes and lamina propria. Activated CD103/β7 binds to CD324 (E-Cadherin) which is involved in the homing of lymphocytes to and their retention at intestinal epithelium.
Development References (4)
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Le Floc'h A, Jalil A, Vergnon I, et al. Alpha E beta 7 integrin interaction with E-cadherin promotes antitumor CTL activity by triggering lytic granule polarization and exocytosis.. J Exp Med. 2007; 204(3):559-70. (Biology). View Reference
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Nie J, Carpenter AC, Chopp LB, et al. The transcription factor LRF promotes integrin β7 expression by and gut homing of CD8αα+ intraepithelial lymphocyte precursors.. Nat Immunol. 2022; 23(4):594-604. (Biology). View Reference
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Qi J, Zhang K, Zhang Q, et al. Identification, characterization, and epitope mapping of human monoclonal antibody J19 that specifically recognizes activated integrin α4β7.. J Biol Chem. 2012; 287(19):15749-59. (Biology). View Reference
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Sato T, Ishihara S, Marui R, Takagi J, Katagiri K. Dissection of α4β7 integrin regulation by Rap1 using novel conformation-specific monoclonal anti-β7 antibodies.. Sci Rep. 2020; 10(1):13221. (Immunogen: Flow cytometry). View Reference
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