Flow cytometry analysis of Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, FITC (31584) was performed using HeLa cells stained with HDAC4 Antibody (711390). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with HDAC4 antibody or with rabbit isotype control at 3-5 ug/million cells in 2.5% BSA and incubated for 2 hours at room temperature. The cells were then labeled with Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, FITC (31584) at a dilution of 1:1000 for 1 hour at room temperature. A representative 10,000 cells were acquired and analyzed for each sample using the Attune® NxT Acoustic Focusing Cytometer. Panels a, b and c represent cells stained with the secondary antibody alone, isotype control and HDAC4 antibody respectively. Median fluorescence intensity from the three samples is compared in panel d.
|Tested species reactivity||Rabbit|
|Host / Isotype||Mouse / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against mouse, human, goat and sheep serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:1000|
|Immunohistochemistry (IHC)||1:50 - 1:200|
|Immunoprecipitation (IP)||1:50 - 1:200|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31584 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31584 reacts with the heavy chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins, but does not react against non-immunoglobulin serum proteins. This antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with mouse, human, goat, and sheep serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Store product protected from light at 4°C until opened. To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C. Fluorescein Amax= 492 nm; Emax= 520 nm. Fluorophore/Protein: 7.6 ug/mg; 2.9 moles FITC per mole IgG. (lot-dependent).
Reconstitute with 0.75 ml of distilled water (1.5 mg/ml after restoration).
Country of Origin: USA
Thermo Scientific Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.