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Multicolor flow cytometric analysis of H-2Kb expression on C57BL/6 mouse splenocytes. Splenic leucocytes from a BALB/c (Left Panel) or a C57BL/6 (Right Panel) mouse were stained with Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram) or with Alexa Fluor® 647 Mouse Anti-Mouse H-2Kb antibody (Cat. No. 562832; solid line histogram). The flow cytometric fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of H-2Kb expression on C57BL/6 mouse splenocytes. Splenic leucocytes from a BALB/c (Left Panel) or a C57BL/6 (Right Panel) mouse were stained with Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram) or with Alexa Fluor® 647 Mouse Anti-Mouse H-2Kb antibody (Cat. No. 562832; solid line histogram). The flow cytometric fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
H2 class I histocompatibility alloantigen Kb
Mouse (QC Testing)
Mouse BALB/c IgG2a, κ
Mouse C57BL/6 Splenocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
14972
AB_2737824
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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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).
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO
Size
100 Tests
Cat No.
557715
562832 Rev. 1
Antibody Details
AF6-88.5
The AF6-88.5 antibody recognizes the H-2Kb MHC class I alloantigen. Reactivity with other haplotypes (e.g., d, f, j, k, p, q, r, s, u, v) has not been observed.
562832 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
562832 Rev.1
Citations & References
Development References (2)
-
Pérarnau B, Saron MF, Reina San Martin B, et al. Single H2Kb, H2Db and double H2KbDb knockout mice: peripheral CD8+ T cell repertoire and anti-lymphocytic choriomeningitis virus cytolytic responses.. Eur J Immunol. 1999; 29(4):1243-52. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Wall KA, Lorber MI, Loken MR, McClatchey S, Fitch FW. Inhibition of proliferation of MIs- and Ia-reactive cloned T cells by a monoclonal antibody against a determinant shared by I-A and I-E.. J Immunol. 1983; 131(3):1056-64. (Clone-specific: Blocking, Flow cytometry, Immunoprecipitation). View Reference
562832 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.
