Immunohistochemistry analysis of SIRT5 showing staining in the cytoplasm of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a SIRT5 Mouse Monoclonal Antibody (730109) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||34-302 of 310 of SIRT5 Protein.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||0.5 mg/ml|
* 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.
|Immunoprecipitation (IP)||See 1 publications below|
SIRT5 is a human member of a family of proteins called Sirtuins (Sir2-like proteins) and are present in prokaryotes and eukaryotes. All Sir2-like proteins have a sirtuin core domain, which contains a series of sequence motifs conserved in organisms ranging from bacteria to humans. Bacterial, yeast and mammalian sirtuins are able to metabolize NAD and possibly at as mono-ADP-ribosyltransferases. The enzymatic function of sirtuins is not yet completely understood but recent reports of histone-activated Sir2-mediated NAD metabolism and NAD-activated Sir2-mediated histone deacetylation suggest a possible coupled reciprocal activation mechanism involving interactions of Sir2 with NAD and the N epsilon-acetyl-lysine groups of acetylated histones.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.
730109 was used in immunoprecipitation to describe a mass spectrometry-based method for scoring immunoprecipitation antibody quality
|Marcon E,Jain H,Bhattacharya A,Guo H,Phanse S,Pu S,Byram G,Collins BC,Dowdell E,Fenner M,Guo X,Hutchinson A,Kennedy JJ,Krastins B,Larsen B,Lin ZY,Lopez MF,Loppnau P,Miersch S,Nguyen T,Olsen JB,Paduch M,Ravichandran M,Seitova A,Vadali G,Vogelsang MS,Whiteaker JR,Zhong G,Zhong N,Zhao L,Aebersold R,Arrowsmith CH,Emili A,Frappier L,Gingras AC,Gstaiger M,Paulovich AG,Koide S,Kossiakoff AA,Sidhu SS,Wodak SJ,Gräslund S,Greenblatt JF,Edwards AM||Nature methods (12:725)||2015|