V500 Hamster IgG2, κ Isotype Control
Clone B81-3 (RUO)
- Brand BD Horizon™
- Alternative Name Anti-KLH
- Concentration 0.2 mg/ml
- Isotype Armenian Hamster IgG2, κ
- Reactivity Mouse, Rat (QC Testing)
Isotype control, Flow cytometry (Routinely Tested)
- Immunogen Keyhole Limpet Hemocyanin
- Storage Buffer Aqueous buffered solution containing protein stabilizer, glycerol and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The B81-3 monoclonal antibody was raised against keyhole limpet hemocyanin (KLH), an antigen not expressed by mammalian cells or cell lines. Intended for use as a hamster immunoglobulin isotype control, the B81-3 antibody can be used as a negative control to help differentiate non-specific background signal when compared against a signal from a specific hamster antibody. The purified B81-3 antibody may also serve as a useful standard for some ELISA applications. KLH is a oxygen carrying metalloprotein from a species of keyhole limpet that lives off the coast of California and is often used as a carrier protein to help elicit immune responses to haptens. Since KLH is phylogenetically distant from many mammalian proteins, it often has very minimal to no cross-reactivity for many immunological assays.
The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is
available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.
When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
BD Horizon™ V500 is a novel organic dye excited by the violet laser. This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan. BD Horizon V500 cannot be used simultaneously with AmCyan or Pacific Orange™.
Suggested Companion Products
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- An isotype control should be used at the same concentration as the antibody of interest.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
- BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- 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.
To minimize non-specific staining via Fc receptors, we recommend the use of Mouse BD Fc Block™, purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).