Description
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.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Format
- Format
BV421
- Excitation Source
Violet 405 nm
- Excitation Max
407 nm
- Emission Max
421 nm
BV421 is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.
Suggested Companion Products
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
