Skip to main content Skip to navigation
Alexa Fluor® 647 Rat Anti-Mouse OX40 Ligand (CD252)
Alexa Fluor® 647 Rat Anti-Mouse OX40 Ligand (CD252)

Flow cytometric analysis of mouse OX40 ligand (CD252) expression on stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated (4 days) in culture with Purified NA/LE Hamster Anti-Mouse CD40 antibody (Cat. No. 553721) and Goat Anti-Mouse IgM (Jackson Immunoresearch 115-006-020). The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and 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 OX40 Ligand (CD252) antibody (Cat. No. 565531, solid line histogram). The fluorescence histogram showing OX40 ligand (CD252) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of mouse OX40 ligand (CD252) expression on stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated (4 days) in culture with Purified NA/LE Hamster Anti-Mouse CD40 antibody (Cat. No. 553721) and Goat Anti-Mouse IgM (Jackson Immunoresearch 115-006-020). The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and 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 OX40 Ligand (CD252) antibody (Cat. No. 565531, solid line histogram). The fluorescence histogram showing OX40 ligand (CD252) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
Down Arrow Up Arrow


BD Pharmingen™
Tnfsf4; OX40 ligand; OX40L; TXGP1; Txgp1l; gp34; Ath-1, Ath1, CD134L
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Rat NRK-52E cell line transfected with mouse OX-40L gene (Txgp1l)
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739283
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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565531 Rev. 2
Antibody Details
Down Arrow Up Arrow
RM134L

The RM134L antibody reacts with CD252 (OX-40 Ligand, OX-40L), a member of the NGF/TNF superfamily which is present on antigen-presenting cells and activated B lymphocytes. OX-40L interacts with OX-40 Antigen (CD134) found predominantly on activated T cells. This ligand-receptor pair is grouped with pairs such as CD40-CD40L and CD80- or CD86-CD28, which contribute significantly to B-cell/T-cell interaction during the immune response. The OX-40L-OX-40 interaction is reciprocally costimulatory in that both T cells and B cells are activated in cross-linking. Stimulation via OX-40 Antigen increases the proliferative and IL-2 production responses of activated T cells, while stimulation via OX-40L enhances proliferation and Ig secretion by activated B cells. The RM134L mAb stains B cells activated for four days with anti-IgM plus anti-CD40 (Clone HM40-3) antibodies. An increased binding of OX-40-Ig fusion protein to mouse splenic B cells was observed when B cells were treated with lipopolysaccharide (LPS), suggesting that OX-40L expression is augmented in LPS-activated splenic B cells when compared to resting cells, but this observation could not be confirmed with the RM134L mAb. In addition, other studies with OX-40-Ig fusion protein detected OX-40L on CD4+ and CD8+ activated splenic T cells, but OX-40L was not detected on T cells with the RM134L mAb. Similar results have been reported by others. The RM134L mAb inhibits the binding of OX-40-Ig fusion protein to OX-40L transfectants and blocks the costimulatory activity of OX-40L.

565531 Rev. 2
Format Details
Down Arrow Up Arrow
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
565531 Rev.2
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. Akiba H, Oshima H, Takeda K, et al. CD28-independent costimulation of T cells by OX40 ligand and CD70 on activated B cells. J Immunol. 1999; 162(12):7058-7066. (Immunogen: Blocking, Flow cytometry, Inhibition, Neutralization). View Reference
  2. Baum PR, Gayle RB 3rd, Ramsdell F, et al. Molecular characterization of murine and human OX40/OX40 ligand systems: identification of a human OX40 ligand as the HTLV-1-regulated protein gp34. EMBO J. 1994; 13(17):3992-4001. (Biology). View Reference
  3. Calderhead DM, Buhlmann JE, van den Eertwegh AJ, Claassen E, Noelle RJ, Fell HP. Cloning of mouse Ox40: a T cell activation marker that may mediate T-B cell interactions. J Immunol. 1993; 151(10):5261-5271. (Biology). View Reference
  4. Godfrey WR, Fagnoni FF, Harara MA, Buck D, Engleman EG. Identification of a human OX-40 ligand, a costimulator of CD4+ T cells with homology to tumor necrosis factor. J Exp Med. 1994 August; 180(2):757-762. (Biology). View Reference
  5. Stuber E, Neurath M, Calderhead D, Fell HP, Strober W. Cross-linking of OX40 ligand, a member of the TNF/NGF cytokine family, induces proliferation and differentiation in murine splenic B cells. Immunity. 1995; 2(5):507-521. (Biology). View Reference
View All (5) View Less
565531 Rev. 2

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.