Hydroxysteroid Dehydrogenase, 11-beta, Type I Enzyme | HSD11B2 enzyme
Hydroxysteroid Dehydrogenase, 11-beta, Type I Mouse (11b-HSD1) Control Peptide
Gene Names
HSD11B2; AME; AME1; HSD2; HSD11K; SDR9C3
Applications
ELISA, Western Blot
Purity
Highly purified
Synonyms
Hydroxysteroid Dehydrogenase; 11-beta; Type I; Type I Mouse (11b-HSD1) Control Peptide; HSD11B2 enzyme
Specificity
Synthetic peptide 16aa mapping near the N-terminus of mouse 11b-HSD1. No significant sequence homology is seen with 11b-HSD2 or other proteins. Species sequence homology
Purity/Purification
Highly purified
Form/Format
Supplied as a liquid in PBS, pH 7.5
Concentration
1mg/ml (varies by lot)
Applicable Applications for HSD11B2 enzyme
ELISA (EL/EIA) and Antibody blocking. Not suitable for use in Western Blot.
Other applications not tested.
Other applications not tested.
Application Notes
ELISA: 50-100ng/well
Antibody Blocking: 5-10ug control peptide per 1ul MBS625504(antiserum)or per 1ug MBS625247 (affintity purified antibody).
Optimal dilutions to be determined by the researcher.
Antibody Blocking -
Principle: It is not uncommon to see more than 1 band in Western Blot when probed with a given antibody or see more diffuse staining in immuno-localization studies. The question arises which one of this band(s)/staining is specific. The antibody specificity is generally studied by competing with excess of antigen (peptide or protein) or immuno-neutralization with the antigens. ln principle, a small volume of antibodv (e.g. 1-5ul) is first reacted with excess peptide (5-50 fold over the antibodv e.g. 1 ug antibody reacted with 5-50ug peptide; exact amounts determined by titration) to neutralize it. The neutralized antibody can no longer subsequently bind to another antigen (a band of interest) or staining pattern. So the band(s)/staining that is competed bv the antigen/peptide is supposedly specific. lf more than one band disappears by peptide/antigen competition then those bands have the antigenic determinants and could be considered either fragments of the large antigen or multimer.
General Protocol for Antibody Blocking:
1. Determine the amount of antibody that is-needed for 1 strip (e.g. 2ml). For example. an antibody has given desired bands at 1:1000 dilution. So you will need 1ul/ml antibody (2ul antibody for 2ml antibody solution). lf an antibody were used at 1:5000 dilution then you would only need 0.2ul/ml (use 2ul of 1:10 dilution for better acturacy).
2. Prepare 2 tubes
2a. Label Tube 1 as "+peptide". Label Tube 2 as "-peptide".
2b. To Tube 1, add antigen/peptide solution (10-50ug peptide or 10-100ul antigen added).
2c. To Tube 2, add same volume PBS (no peptide/antigen) to the other tube. Mix gently.
3. Incubate both tubes at 37°C for 1-2 hrs and 2-24 hrs at 4°C.
4. Centrifuge the tubes for 15 min at 4°C in a microfuge (10-15000rpm) to pellet any immune complexes. Carefully remove the supernatant. lf no visible pellet is seen, then just leave approx. 5-l0ul at the bottom to avoid disturbing invisible immune complexes. If you do not centrifuge the solution, it may give high background.
5. After centrifugation, make up the volume of supernatant to 2ml (or what is necessary depending upon the initial antibody taken) with buffer (PBS, Tween with or without BSA or milk or any buffer that was used initially). Use both antibodies (with and without antigen/peptide) for Western Blot or immuno-localization.
6. Observe the bands/staining that disappear.
Notes:
1. The greater the antibody titer or initial volume of antibody taken, the greater will be the antigen/peptide necessary to completely block the antibody activity.
2. A partial inhibition of antibody activity is an indication that more antigen/peptide will be needed to completely block the antibody.
3. It may be necessary to optimize the amount of antigen/peptides by adding various amounts to a fixed concentration of antibody.
Antibody Blocking: 5-10ug control peptide per 1ul MBS625504(antiserum)or per 1ug MBS625247 (affintity purified antibody).
Optimal dilutions to be determined by the researcher.
Antibody Blocking -
Principle: It is not uncommon to see more than 1 band in Western Blot when probed with a given antibody or see more diffuse staining in immuno-localization studies. The question arises which one of this band(s)/staining is specific. The antibody specificity is generally studied by competing with excess of antigen (peptide or protein) or immuno-neutralization with the antigens. ln principle, a small volume of antibodv (e.g. 1-5ul) is first reacted with excess peptide (5-50 fold over the antibodv e.g. 1 ug antibody reacted with 5-50ug peptide; exact amounts determined by titration) to neutralize it. The neutralized antibody can no longer subsequently bind to another antigen (a band of interest) or staining pattern. So the band(s)/staining that is competed bv the antigen/peptide is supposedly specific. lf more than one band disappears by peptide/antigen competition then those bands have the antigenic determinants and could be considered either fragments of the large antigen or multimer.
General Protocol for Antibody Blocking:
1. Determine the amount of antibody that is-needed for 1 strip (e.g. 2ml). For example. an antibody has given desired bands at 1:1000 dilution. So you will need 1ul/ml antibody (2ul antibody for 2ml antibody solution). lf an antibody were used at 1:5000 dilution then you would only need 0.2ul/ml (use 2ul of 1:10 dilution for better acturacy).
2. Prepare 2 tubes
2a. Label Tube 1 as "+peptide". Label Tube 2 as "-peptide".
2b. To Tube 1, add antigen/peptide solution (10-50ug peptide or 10-100ul antigen added).
2c. To Tube 2, add same volume PBS (no peptide/antigen) to the other tube. Mix gently.
3. Incubate both tubes at 37°C for 1-2 hrs and 2-24 hrs at 4°C.
4. Centrifuge the tubes for 15 min at 4°C in a microfuge (10-15000rpm) to pellet any immune complexes. Carefully remove the supernatant. lf no visible pellet is seen, then just leave approx. 5-l0ul at the bottom to avoid disturbing invisible immune complexes. If you do not centrifuge the solution, it may give high background.
5. After centrifugation, make up the volume of supernatant to 2ml (or what is necessary depending upon the initial antibody taken) with buffer (PBS, Tween with or without BSA or milk or any buffer that was used initially). Use both antibodies (with and without antigen/peptide) for Western Blot or immuno-localization.
6. Observe the bands/staining that disappear.
Notes:
1. The greater the antibody titer or initial volume of antibody taken, the greater will be the antigen/peptide necessary to completely block the antibody activity.
2. A partial inhibition of antibody activity is an indication that more antigen/peptide will be needed to completely block the antibody.
3. It may be necessary to optimize the amount of antigen/peptides by adding various amounts to a fixed concentration of antibody.
Source
Mouse
Species sequence homology
100% conserved in the mouse and rat, 87% in sheep and 81% in human and chicken, 75% in porcine and 68% in rabbit 11b-HSD1. No significant sequence homology is seen with other proteins. Control peptide for MBS625504(antiserum)or per 1ug MBS625247 (affintity purified antibody)
Preparation and Storage
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt 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.
Related Product Information for HSD11B2 enzyme
Synthetic peptide corresponding to 16aa of mouse 11b-HSD1 conjugated to KLH.
11-Beta-hydroxysteroid dehydrogenase (11b-HSD) is a microsomal short chain dehydrogenase/reductase (SDR) which catalyzes the inter-conversion of biologically active glucocorticoid (cortisol in human and corticosterone in rats and mice) and inactive glucocorticoid (cortisone and 11-dehydrocorticosterone). Two tissue specific isoforms (11b-HSD1 and 11b-HSD2) of 11b-HSD with two different functions regarding glucocorticoid availability, have been identified. The decreased 11-beta-hydroxy oxidation of cortisol results in Apparent Mineralocorticoid Excess (AME) disorder which is manifested by hypertension, hypokalemia, low plasma renin activity, and responsiveness to spironolactone. AME is principally a disorder of juveniles and children with this condition oxidize cortisol to cortisone poorly but carry out the reverse process unimpaired. AME arises from mutations in the 11-beta-HSD2 gene. The glucocorticoids can be produced locally by 11beta-HSD1 and increased visceral accumulation of glucocorticoids results in visceral obesity, insulin resistant diabetes, hyperlipidemia and hyperphagia.
11betaHSD-1 (variously termed as HSD11L; mouse, 292 aa, rat 287 aa, human 292 aa) is a ~35kD glycosylated membrane-protein, oriented into the lumen of endoplasmic reticulum. This isoform is the sole 11b-reductase in the body and exerts two separate enzymatic activities: 11-beta-dehydrogenase (cortisol to cortisone) and 11-oxoreductase (cortisone to cortisol) in vitro. In vivo, it acts mainly as reductase producing active cortisol. The enzyme also plays an important role in xenobiotic carbonyl compound detoxification processes. 11b-HSD1 is expressed in a wide array of tissues, with highest level in Liver and adipose tissues. The increased adipocyte 11b-HSD1 is a common mechanism for visceral obesity and metabolic syndrome. Although the deficiency in 11b-HSD1 activity is not related to AME, it results in a syndrome characterized by an increased adrenocorticotropic hormone (ACTH)-driven androgen production. In mouse, the over-all aa sequence of 11b-HSD1 is approximately 18% identical to that of 11b-HSD2.
11-Beta-hydroxysteroid dehydrogenase (11b-HSD) is a microsomal short chain dehydrogenase/reductase (SDR) which catalyzes the inter-conversion of biologically active glucocorticoid (cortisol in human and corticosterone in rats and mice) and inactive glucocorticoid (cortisone and 11-dehydrocorticosterone). Two tissue specific isoforms (11b-HSD1 and 11b-HSD2) of 11b-HSD with two different functions regarding glucocorticoid availability, have been identified. The decreased 11-beta-hydroxy oxidation of cortisol results in Apparent Mineralocorticoid Excess (AME) disorder which is manifested by hypertension, hypokalemia, low plasma renin activity, and responsiveness to spironolactone. AME is principally a disorder of juveniles and children with this condition oxidize cortisol to cortisone poorly but carry out the reverse process unimpaired. AME arises from mutations in the 11-beta-HSD2 gene. The glucocorticoids can be produced locally by 11beta-HSD1 and increased visceral accumulation of glucocorticoids results in visceral obesity, insulin resistant diabetes, hyperlipidemia and hyperphagia.
11betaHSD-1 (variously termed as HSD11L; mouse, 292 aa, rat 287 aa, human 292 aa) is a ~35kD glycosylated membrane-protein, oriented into the lumen of endoplasmic reticulum. This isoform is the sole 11b-reductase in the body and exerts two separate enzymatic activities: 11-beta-dehydrogenase (cortisol to cortisone) and 11-oxoreductase (cortisone to cortisol) in vitro. In vivo, it acts mainly as reductase producing active cortisol. The enzyme also plays an important role in xenobiotic carbonyl compound detoxification processes. 11b-HSD1 is expressed in a wide array of tissues, with highest level in Liver and adipose tissues. The increased adipocyte 11b-HSD1 is a common mechanism for visceral obesity and metabolic syndrome. Although the deficiency in 11b-HSD1 activity is not related to AME, it results in a syndrome characterized by an increased adrenocorticotropic hormone (ACTH)-driven androgen production. In mouse, the over-all aa sequence of 11b-HSD1 is approximately 18% identical to that of 11b-HSD2.
Product Categories/Family for HSD11B2 enzyme
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
< 3kD
NCBI Official Full Name
Hydroxysteroid (11-beta) dehydrogenase 2
NCBI Official Synonym Full Names
hydroxysteroid (11-beta) dehydrogenase 2
NCBI Official Symbol
HSD11B2
NCBI Official Synonym Symbols
AME; AME1; HSD2; HSD11K; SDR9C3
NCBI Protein Information
corticosteroid 11-beta-dehydrogenase isozyme 2; 11-DH2; 11-beta-HSD2; -HSD11 type II; 11-beta-hydroxysteroid dehydrogenase type 2; 11-beta-hydroxysteroid dehydrogenase type II; NAD-dependent 11-beta-hydroxysteroid dehydrogenase; short chain dehydrogenase/reductase family 9C member 3
UniProt Protein Name
Corticosteroid 11-beta-dehydrogenase isozyme 2
Protein Family
UniProt Gene Name
HSD11B2
UniProt Synonym Gene Names
HSD11K; 11-DH2; 11-beta-HSD2; -HSD11 type II; 11-beta-HSD
UniProt Entry Name
DHI2_HUMAN
NCBI Description
There are at least two isozymes of the corticosteroid 11-beta-dehydrogenase, a microsomal enzyme complex responsible for the interconversion of cortisol and cortisone. The type I isozyme has both 11-beta-dehydrogenase (cortisol to cortisone) and 11-oxoreductase (cortisone to cortisol) activities. The type II isozyme, encoded by this gene, has only 11-beta-dehydrogenase activity. In aldosterone-selective epithelial tissues such as the kidney, the type II isozyme catalyzes the glucocorticoid cortisol to the inactive metabolite cortisone, thus preventing illicit activation of the mineralocorticoid receptor. In tissues that do not express the mineralocorticoid receptor, such as the placenta and testis, it protects cells from the growth-inhibiting and/or pro-apoptotic effects of cortisol, particularly during embryonic development. Mutations in this gene cause the syndrome of apparent mineralocorticoid excess and hypertension. [provided by RefSeq, Feb 2010]
Uniprot Description
Function: Catalyzes the conversion of cortisol to the inactive metabolite cortisone. Modulates intracellular glucocorticoid levels, thus protecting the nonselective mineralocorticoid receptor from occupation by glucocorticoids.
Catalytic activity: An 11-beta-hydroxysteroid + NAD+ = an 11-oxosteroid + NADH.
Enzyme regulation: Inhibited by glycyrrhetinic acid (derived from liquorice), carbenoloxone and 11-alpha-OH-progesterone
By similarity.
Subunit structure: Interacts with ligand-free cytoplasmic NR3C2. Ref.12
Subcellular location: Microsome. Endoplasmic reticulum Ref.24.
Tissue specificity: Found in placenta, kidney, pancreas, prostate, ovary, small intestine and colon.
Involvement in disease: Apparent mineralocorticoid excess (AME) [MIM:218030]: An autosomal recessive form of low-renin hypertension. It is usually diagnosed within the first years of life and is characterized by polyuria and polydipsia, failure to thrive, hypernatremia, severe hypertension with low renin and aldosterone levels, profound hypokalemia with metabolic alkalosis, and most often nephrocalcinosis.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.10 Ref.13 Ref.14 Ref.15 Ref.16 Ref.17 Ref.18 Ref.20 Ref.21 Ref.22 Ref.23 Ref.24
Miscellaneous: Consumption of large amounts of liquorice can lead to apparent mineralocorticoid excess and hypertension.
Sequence similarities: Belongs to the short-chain dehydrogenases/reductases (SDR) family.
Biophysicochemical propertiesKinetic parameters:X.KM=26.1 nM for cortisol (Ref.23) Ref.23 Ref.24KM=785 nM for cortisol (Ref.24)KM=77 nM for cortisteroneVmax=64.1 nmol/h/mg enzyme toward cortisteroneVmax=66 nmol/h/mg enzyme toward cortisol
Research Articles on HSD11B2
Similar Products
Product Notes
The HSD11B2 hsd11b2 (Catalog #AAA653798) is an Enzyme and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Hydroxysteroid Dehydrogenase, 11-beta, Type I can be used in a range of immunoassay formats including, but not limited to, ELISA (EL/EIA) and Antibody blocking. Not suitable for use in Western Blot. Other applications not tested. ELISA: 50-100ng/well Antibody Blocking: 5-10ug control peptide per 1ul MBS625504(antiserum)or per 1ug MBS625247 (affintity purified antibody). Optimal dilutions to be determined by the researcher. Antibody Blocking - Principle: It is not uncommon to see more than 1 band in Western Blot when probed with a given antibody or see more diffuse staining in immuno-localization studies. The question arises which one of this band(s)/staining is specific. The antibody specificity is generally studied by competing with excess of antigen (peptide or protein) or immuno-neutralization with the antigens. ln principle, a small volume of antibodv (e.g. 1-5ul) is first reacted with excess peptide (5-50 fold over the antibodv e.g. 1 ug antibody reacted with 5-50ug peptide; exact amounts determined by titration) to neutralize it. The neutralized antibody can no longer subsequently bind to another antigen (a band of interest) or staining pattern. So the band(s)/staining that is competed bv the antigen/peptide is supposedly specific. lf more than one band disappears by peptide/antigen competition then those bands have the antigenic determinants and could be considered either fragments of the large antigen or multimer. General Protocol for Antibody Blocking: 1. Determine the amount of antibody that is-needed for 1 strip (e.g. 2ml). For example. an antibody has given desired bands at 1:1000 dilution. So you will need 1ul/ml antibody (2ul antibody for 2ml antibody solution). lf an antibody were used at 1:5000 dilution then you would only need 0.2ul/ml (use 2ul of 1:10 dilution for better acturacy). 2. Prepare 2 tubes 2a. Label Tube 1 as "+peptide". Label Tube 2 as "-peptide". 2b. To Tube 1, add antigen/peptide solution (10-50ug peptide or 10-100ul antigen added). 2c. To Tube 2, add same volume PBS (no peptide/antigen) to the other tube. Mix gently. 3. Incubate both tubes at 37°C for 1-2 hrs and 2-24 hrs at 4°C. 4. Centrifuge the tubes for 15 min at 4°C in a microfuge (10-15000rpm) to pellet any immune complexes. Carefully remove the supernatant. lf no visible pellet is seen, then just leave approx. 5-l0ul at the bottom to avoid disturbing invisible immune complexes. If you do not centrifuge the solution, it may give high background. 5. After centrifugation, make up the volume of supernatant to 2ml (or what is necessary depending upon the initial antibody taken) with buffer (PBS, Tween with or without BSA or milk or any buffer that was used initially). Use both antibodies (with and without antigen/peptide) for Western Blot or immuno-localization. 6. Observe the bands/staining that disappear. Notes: 1. The greater the antibody titer or initial volume of antibody taken, the greater will be the antigen/peptide necessary to completely block the antibody activity. 2. A partial inhibition of antibody activity is an indication that more antigen/peptide will be needed to completely block the antibody. 3. It may be necessary to optimize the amount of antigen/peptides by adding various amounts to a fixed concentration of antibody. Researchers should empirically determine the suitability of the HSD11B2 hsd11b2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Hydroxysteroid Dehydrogenase, 11-beta, Type I, Enzyme" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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