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Anti -HDAC5 (HD5, FLJ90614, KIAA0600, NY-CO-9, Histone Deacetylase 5)
Anti -HDAC5 (HD5, FLJ90614, KIAA0600, NY-CO-9, Histone Deacetylase 5)
Pricing
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| Clone |
Host |
Grade |
Applications |
| Polyclonal |
Rabbit |
Affinity Purified |
B |
|
| Catalog # | H1827-65 | | Applications | Suitable for use in Western Blot. Other applications not tested. | | Recommended Dilution | Western 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 Control | HeLa 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 Stability | May 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, 1 | 5,000 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). | | CAS Number | n/a | | Clone Type | Polyclonal | | Isotype | IgG | | Host | Rabbit | | Source | Human | | Concentration | ~1mg/ml | | Form | Supplied 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%. | | Purity | Purified by Protein A affinity chromatography. | | Immunogen | KLH-conjugated synthetic peptides corresponding to amino acids 536-545 and 194-206 (CTKTGELPRQP and C-KSKEPTPGGLNHS) of human HDAC5. | | Specificity | Recognizes HDAC5, Mr 160kD. Species Crossreactivity: Human. Weak crossreactivity with rat and mouse. | | | Important Note | This 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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