Description
RP1-Site-2 Protease is a polyclonal antibody made to the membrane-spanning metalloprotease Site-2 protease. The antibody is made to a synthetic peptide based on the first luminal domain of human site-2 protease. The antibody has been peptide-affinity purified, concentrated to 1.0 mg/ml, with the addition of 0.05% sodium azide as preservative and 50% glycerol as cryoprotectant.
Use
Site-2 protease, also known as S2P, Membrane-bound transcription factor site-2 protease, MBTP2, S2P endopeptidase and sterol-regulatory element-binding proteins intermediate protease, is a metalloprotease of the MM clan, characterized by having the zinc-binding HExxH motif located within a transmembrane domain, and the third binding ligand an aspartic acid locatred in a LDG motif in another transmembrane domain near the carboxyterminal end. Site-2 protease is in the M50A family of the MM clan, and is located in the endoplasmic reticulum membrane. Site-1 protease cleaves SREBPs in the luminal segment, at a leucine-serine sequence in the luminal loop. This cleavage requires binding of SCAP to the SREBPs in the ER. The S1P cleavage allows the site-2 protease to cleave the SREBPs at a leucine-cysteine bond in the following transmembrane segment, releasing the aminoterminal end which acts as a transcription factor. The cleaved aminoterminal fragment acts to control cholesterol synthesis and processing. An arginine-proline sequence 11 residues upstream from the leu-cys cleavage site is important for binding the SREBPs and cleavage of the leu-cys bond. The S1P activity is also regulated by sterol binding, and although S2P is not directly regulated by sterols the S2P cleavage only occurs in sterol-depleted cells. The amino and carboxyterminal residues of S2P are exposed to the cytoplasm. The first transmembrane domain follows the aminotermoinal cytoplasmic sequence, followed by a luminal stretch, a dip back into the membrane, another luminal stretch containing a serine-rich region, an extended transmembrane domain containing the HExxH zinc-binding site, another luminal domain containing a cysteine-rich stretch, another transmembrane domain containing the LDG motif, a brief pass through the lumen, and then a final transmembrane domain followed by the cytoplasmic cap. Full length S2P is a 519 amino acid protein with a predicted mass of 57.4 kDa and a pI of 7.6. A 330 amino acid form is reported, with predicted mass of 36.4 and a pI of 5.7, and a 223 residue form with predicted mass of 24.8 and a pI of 9.3. Both shorter forms are truncated at the carboxyterminal end, and lack the catalytic aspartic acid, thus are thought to be catalytically inactive, if produced. A recommended starting concentration for Western blots is 1:1,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species. FOR RESEARCH USE ONLY; NOT FOR USE IN HUMANS.
Storage
The undiluted antibody solution is stable for approximately 12 months at -20C.
Description
RP2-Site-2 Protease is a polyclonal antibody made to the membrane-spanning metalloprotease Site-2 protease. The antibody is made to a synthetic peptide based on the metalloprotease domain of human site-2 protease. The antibody has been peptide-affinity purified, concentrated to 1.0 mg/ml, with the addition of 0.05% sodium azide as preservative and 50% glycerol as cryoprotectant.
Use
Site-2 protease, also known as S2P, Membrane-bound transcription factor site-2 protease, MBTP2, S2P endopeptidase and sterol-regulatory element-binding proteins intermediate protease, is a metalloprotease of the MM clan, characterized by having the zinc-binding HExxH motif located within a transmembrane domain, and the third binding ligand an aspartic acid locatred in a LDG motif in another transmembrane domain near the carboxyterminal end. Site-2 protease is in the M50A family of the MM clan, and is located in the endoplasmic reticulum membrane. Site-1 protease cleaves SREBPs in the luminal segment, at a leucine-serine sequence in the luminal loop. This cleavage requires binding of SCAP to the SREBPs in the ER. The S1P cleavage allows the site-2 protease to cleave the SREBPs at a leucine-cysteine bond in the following transmembrane segment, releasing the aminoterminal end which acts as a transcription factor. The cleaved aminoterminal fragment acts to control cholesterol synthesis and processing. An arginine-proline sequence 11 residues upstream from the leu-cys cleavage site is important for binding the SREBPs and cleavage of the leu-cys bond. The S1P activity is also regulated by sterol binding, and although S2P is not directly regulated by sterols the S2P cleavage only occurs in sterol-depleted cells. The amino and carboxyterminal residues of S2P are exposed to the cytoplasm. The first transmembrane domain follows the aminotermoinal cytoplasmic sequence, followed by a luminal stretch, a dip back into the membrane, another luminal stretch containing a serine-rich region, an extended transmembrane domain containing the HExxH zinc-binding site, another luminal domain containing a cysteine-rich stretch, another transmembrane domain containing the LDG motif, a brief pass through the lumen, and then a final transmembrane domain followed by the cytoplasmic cap. Full length S2P is a 519 amino acid protein with a predicted mass of 57.4 kDa and a pI of 7.6. A 330 amino acid form is reported, with predicted mass of 36.4 and a pI of 5.7, and a 223 residue form with predicted mass of 24.8 and a pI of 9.3. Both shorter forms are truncated at the carboxyterminal end, and lack the catalytic aspartic acid, thus are thought to be catalytically inactive, if produced. A recommended starting concentration for Western blots is 1:1,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species. FOR RESEARCH USE ONLY; NOT FOR USE IN HUMANS.
Storage
The undiluted antibody solution is stable for approximately 12 months at -20C.
Description
RP3-Site-2 Protease is a polyclonal antibody made to the membrane-spanning metalloprotease Site-2 protease. The antibody is made to a synthetic peptide based on the cysteine-rich luminal domain of human site-2 protease. The antibody has been peptide-affinity purified, concentrated to 1.0 mg/ml, with the addition of 0.05% sodium azide as preservative and 50% glycerol as cryoprotectant.
Use
Site-2 protease, also known as S2P, Membrane-bound transcription factor site-2 protease, MBTP2, S2P endopeptidase and sterol-regulatory element-binding proteins intermediate protease, is a metalloprotease of the MM clan, characterized by having the zinc-binding HExxH motif located within a transmembrane domain, and the third binding ligand an aspartic acid locatred in a LDG motif in another transmembrane domain near the carboxyterminal end. Site-2 protease is in the M50A family of the MM clan, and is located in the endoplasmic reticulum membrane. Site-1 protease cleaves SREBPs in the luminal segment, at a leucine-serine sequence in the luminal loop. This cleavage requires binding of SCAP to the SREBPs in the ER. The S1P cleavage allows the site-2 protease to cleave the SREBPs at a leucine-cysteine bond in the following transmembrane segment, releasing the aminoterminal end which acts as a transcription factor. The cleaved aminoterminal fragment acts to control cholesterol synthesis and processing. An arginine-proline sequence 11 residues upstream from the leu-cys cleavage site is important for binding the SREBPs and cleavage of the leu-cys bond. The S1P activity is also regulated by sterol binding, and although S2P is not directly regulated by sterols the S2P cleavage only occurs in sterol-depleted cells. The amino and carboxyterminal residues of S2P are exposed to the cytoplasm. The first transmembrane domain follows the aminotermoinal cytoplasmic sequence, followed by a luminal stretch, a dip back into the membrane, another luminal stretch containing a serine-rich region, an extended transmembrane domain containing the HExxH zinc-binding site, another luminal domain containing a cysteine-rich stretch, another transmembrane domain containing the LDG motif, a brief pass through the lumen, and then a final transmembrane domain followed by the cytoplasmic cap. Full length S2P is a 519 amino acid protein with a predicted mass of 57.4 kDa and a pI of 7.6. A 330 amino acid form is reported, with predicted mass of 36.4 and a pI of 5.7, and a 223 residue form with predicted mass of 24.8 and a pI of 9.3. Both shorter forms are truncated at the carboxyterminal end, and lack the catalytic aspartic acid, thus are thought to be catalytically inactive, if produced. A recommended starting concentration for Western blots is 1:1,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species. FOR RESEARCH USE ONLY; NOT FOR USE IN HUMANS.
Storage
The undiluted antibody solution is stable for approximately 12 months at -20C.
