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|Antigen||BCL2 Binding Component 3 (BBC3) Antibodies|
|Epitope||C-Term, AA 150-193 Alternatives|
|Conjugate||This BBC3 antibody is un-conjugated Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blotting (WB)
|2 references available|
|Supplier||Log in to see|
Product Details anti-BBC3 AntibodyTarget Details BBC3 Application Details Handling References for anti-BBC3 antibody (ABIN153058) Images
|Purification||Immunogen affinity purified|
|Immunogen||A synthetic peptide made to a region within the C-terminus of the human PUMA protein sequence (between residues 150-193). [UniProt# Q9BXH1]|
Target Details BBC3Product Details anti-BBC3 Antibody Application Details Handling References for anti-BBC3 antibody (ABIN153058) Images back to top
|Alternative Name||PUMA (BBC3 Antibody Abstract)|
|Background||Gene Symbol: BBC3|
|Molecular Weight||Theoretical MW: 16 kDa|
|Research Area||Metabolism, Apoptosis/Necrosis, Cancer|
|Pathways||p53 Signaling, Positive Regulation of Endopeptidase Activity|
Application DetailsProduct Details anti-BBC3 Antibody Target Details BBC3 Handling References for anti-BBC3 antibody (ABIN153058) Images back to top
|Application Notes||Western Blot 2-4 μg/mL, Immunocytochemistry/Immunofluorescence 1:20-1:100This PUMA antibody is useful for Western blot and Immunocytochemistry/Immunofluorescence. By WB a band at ~16 kDa is seen, representing the beta form of PUMA. Reactivity against the alpha form of PUMA is unknown. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Protocol specific for PUMA Antibody Western Blot Protocol
1. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 35 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 0.5 % BSA in TBS for 1 hour.
. Dilute the rabbit anti-PUMA primary antibody in blocking buffer and incubate 2 hours at room temperature.
. Wash the membrane in water for 5 minutes and apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
. Wash the blot in TBS containing 0.05-0.1 % Tween-20 for 10-20 minutes.
. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturer's instructions (Amersham's ECL is the standard reagent used).Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.
|Restrictions||For Research Use only|
HandlingProduct Details anti-BBC3 Antibody Target Details BBC3 Application Details References for anti-BBC3 antibody (ABIN153058) Images back to top
Tris-Glycine and 0.15M NaCl
Buffer contains: 0.05 % Sodium Azide
|Precaution of Use||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Handling Advice||Avoid freeze-thaw cycles|
|Storage||4 °C,-20 °C|
|Storage Comment||Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.|
References for anti-BBC3 antibody (ABIN153058)Product Details anti-BBC3 Antibody Target Details BBC3 Application Details Handling Images back to top
|Product cited in:||
Stylianou, Clarke, Brennan: "Aberrant activation of notch signaling in human breast cancer." in: Cancer research, Vol. 66, Issue 3, pp. 1517-25, 2006 (Sample species: Human). Further details: Western Blotting
Yu, Wang, Kinzler, Vogelstein, Zhang: "PUMA mediates the apoptotic response to p53 in colorectal cancer cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, Issue 4, pp. 1931-6, 2003
ImagesProduct Details anti-BBC3 Antibody Target Details BBC3 Application Details Handling References for anti-BBC3 antibody (ABIN153058) back to top