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You are here:Home » Antibodies » Abs to Enzymes, Deacetylase » Anti -HDAC5 (HD5, FLJ90614, KIAA0600, NY-CO-9, Histone Deacetylase 5)

Anti -HDAC5 (HD5, FLJ90614, KIAA0600, NY-CO-9, Histone Deacetylase 5)

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Specifications

Clone Host Grade Applications
Polyclonal Rabbit Affinity Purified B
Catalog #H1827-65
ApplicationsSuitable for use in Western Blot. Other applications not tested.
Recommended DilutionWestern Blot: 1-4ug/ul detects HDAC5 from a HeLa nuclear extract. HeLa nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HDAC5 (1ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Optimal dilutions to be determined by the researcher.
Positive Antigen ControlHeLa nuclear extract. Add an equal volume of Laemmli reducing sample buffer to 10ul of extract and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunoblot Protocol 1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a nuclear extract and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. 2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk (PBS-MLK) for 60 minutes at 20-25°C with constant agitation. 3. Incubate the nitrocellulose with 1-4ug/ml of anti-HDAC5, diluted in freshly prepared PBS-MLK overnight with agitation at 4°C. 4. Wash the nitrocellulose twice with water. 5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-rabbit HRP conjugated IgG, 15,000 dilution was used) in PBS-MLK for 1.5 hours at room temperature with agitation. 6. Wash the nitrocellulose with water twice. 7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 3-5 minutes. 8. Rinse the nitrocellulose in 4-5 changes of water.
9. Use detection method of choice (enhanced chemiluminescence was used).
Clone TypePolyclonal
IsotypeIgG
HostRabbit
SourceHuman
Concentration~1mg/ml
FormSupplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide before the addition of glycerol to 30%.
PurityPurified by Protein A affinity chromatography.
ImmunogenKLH-conjugated synthetic peptides corresponding to amino acids 536-545 and 194-206 (CTKTGELPRQP and C-KSKEPTPGGLNHS) of human HDAC5.
SpecificityRecognizes HDAC5, Mr 160kD. Species Crossreactivity: Human. Weak crossreactivity with rat and mouse.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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