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Direct ELISA analysis of Histone H4 was performed by coating wells of a 96-well plate with 100ul per well of recombinant Histone H4 or Histone H3 protein diluted in carbonate/bicarbonate buffer (Product # 28382), at a concentration of 1ug/ml. Wells of the plate were washed, blocked with Superblock blocking buffer (Product # 37536), and incubated with 100ul per well of a mouse anti-Histone H4 monoclonal antibody (Product # MA3-050) starting at a concentration of 1ug/ml and serially diluting 2-fold to a concentration of 2ng/ml for 1 hour at 37C. The plate was washed, then incubated with 100ul per well of a Biotin-conjugated goat anti-mouse IgG secondary antibody (Product # 31807) at a dilution of 1:5000 for 30 minutes at 37C followed by washing and incubated with HRP-conjugated streptavidin (Product # 21126) at a dilution of 1:5000 for 30 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1:20,000 - 1:400,000|
|Flow Cytometry (Flow)||1:200 - 1:1,000|
|Immunocytochemistry (ICC)||1:500 - 1:5,000|
|Immunofluorescence (IF)||1:500 - 1:5,000|
|Immunohistochemistry (IHC)||1:500 - 1:5,000|
|Immunoprecipitation (IP)||1:500 - 1:5,000|
|Western Blot (WB)||1:20,000 - 1:400,000|
* 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 # 31807 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31807 reacts with the heavy chains of mouse IgG and IgM and with the light chains common to most mouse immunoglobulins, but does not react with non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Store product 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.
Reconstitute with 2.0 ml of distilled water (1.2 mg/ml after restoration).
Country of Origin: USA
Thermo Scientific Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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.