Flow cytometric analysis of CD49d expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat Anti-Mouse CD49d antibody (Cat. No. 564394; solid line histogram). The fluorescence histogram showing CD49d expression (or Ig Isotype control staining) was 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.
Flow cytometric analysis of CD49d expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat Anti-Mouse CD49d antibody (Cat. No. 564394; solid line histogram). The fluorescence histogram showing CD49d expression (or Ig Isotype control staining) was 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.
Product Details
BD Pharmingen™
Integrin α4; Itga4; Integrin alpha-4; LPAM alpha; VLA-4 alpha; VLA-4a
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2b, κ
AKR/Cum mouse spontaneous T lymphoma line TK1
Flow cytometry (Routinely Tested)
0.2 mg/ml
16401
AB_2738787
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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Rat IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557691
564394 Rev. 1
Antibody Details
R1-2
The R1-2 monoclonal antibody specifically binds to α4 chain (CD49d), which is expressed as a heterdimer with either of two β, β1 or β7 (also known as βp). The α4β1 integrin (VLA-4, CD49d/CD29) is expressed on most peripheral lymphocytes, thymocytes, and monocytes; while the α4β7 integrin (LPAM-1) is expressed on peripheral lymphocytes, but on only a small subset of thymocytes. These integrins mediate a variety of cell-cell and cell-matrix interactions, recognizing the ligands VCAM-1 (CD106) and fibronectin. There is evidence that levels of VLA-4 expression regulate the transendothelial migration of T lymphocytes into inflamed tissues. Integrin α4β7 also preferentially binds to the mucosal vascular addressin, MAdCAM-1. The R1-2 antibody blocks some α4 integrin-mediated binding functions. In combination with mAb 9C10 (MFR4.B) (Cat. No. 553313), binding of VLA-4 expressing cells to VCAM-1 can be almost completely inhibited.
564394 Rev. 1
Format Details
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
564394 Rev.1
Citations & References
Development References (8)
-
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. (Biology). 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. (Biology). View Reference
-
Ferguson TA, Kupper TS. Antigen-independent processes in antigen-specific immunity. A role for alpha 4 integrin. J Immunol. 1993; 150(4):1172-1182. (Biology). View Reference
-
Holzmann B, McIntyre BW, Weissman IL. Identification of a murine Peyer's patch--specific lymphocyte homing receptor as an integrin molecule with an alpha chain homologous to human VLA-4 alpha. Cell. 1989; 56(1):37-46. (Immunogen). View Reference
-
Holzmann B, Weissman IL. Peyer's patch-specific lymphocyte homing receptors consist of a VLA-4-like alpha chain associated with either of two integrin beta chains, one of which is novel. EMBO J. 1989; 8(6):1735-1741. (Clone-specific: Immunoprecipitation, Inhibition). View Reference
-
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
-
Kinashi T, Springer TA. Adhesion molecules in hematopoietic cells. Blood Cells. 1994; 20(1):25-44. (Biology). View Reference
-
Romanic AM, Graesser D, Baron JL, Visintin I, Janeway CA Jr, Madri JA. T cell adhesion to endothelial cells and extracellular matrix is modulated upon transendothelial cell migration. Lab Invest. 1997; 76(1):11-23. (Biology). View Reference
564394 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.
