FITC Mouse IgG2b, κ Isotype Control
Clone 27-35 (RUO)
- Brand BD Pharmingen™
- Alternative Name Anti-Dansyl; Isotype control (anti-Dansyl)
- Concentration 0.5 mg/ml
- Isotype Mouse C.SW IgG2b, κ
Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The mouse IgG2b, κ immunoglobulin isotype control monoclonal antibody 27-35 is specific for the hapten dansyl (5-[dimethylamino] naphthalene-1-sulfonyl). This hapten is not expressed on human cells or human cell lines. The 27-35 immunoglobulin was selected as an isotype control following testing which demonstrated low background staining on a variety of mouse and human tissues.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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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 with FITC under optimum conditions, and unreacted FITC 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.
- 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.
Immunofluorescent Staining and Flow Cytometic Analysis: The PE-and FITC-conjugated 27-35 immunoglobulin (Cat. No. 555057, 555058 respectively) are suitable mouse IgG2b isotype controls for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized rat or human cells for flow cytometric analysis. Use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells) (see image, middle panel). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit our web site, www.bdbiosciences.com/us/s/resources and refer to the protocols section under Intracellular Flow.