Alexa Fluor® 647 Rat Anti-Mouse OX40 Ligand (CD252)
Clone RM134L (RUO)
- Brand BD Pharmingen™
- Alternative Name Tnfsf4; OX40 ligand; OX40L; TXGP1; Txgp1l; gp34; Ath-1, Ath1, CD134L
- Concentration 0.2 mg/ml
- Isotype Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen Rat NRK-52E cell line transfected with mouse OX-40L gene (Txgp1l)
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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.
- 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.
- 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.
- 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).
- 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/pharmingen/protocols for technical protocols.