protein

Hydroxysteroid Dehydrogenase, 11-beta, Type I Mouse (11b-HSD1) Control Peptide

MBS653798

0.1 mg

395 USD

Enzyme

Hydroxysteroid Dehydrogenase, 11-beta, Type I Mouse (11b-HSD1) Control Peptide

[Hydroxysteroid Dehydrogenase, 11-beta, Type I]

[Hydroxysteroid (11-beta) dehydrogenase 2; Corticosteroid 11-beta-dehydrogenase isozyme 2; 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; hydroxysteroid (11-beta) dehydrogenase 2; 11-beta-hydroxysteroid dehydrogenase type 2; 11-DH2; 11-beta-HSD2; 11-beta-hydroxysteroid dehydrogenase type II; -HSD11 type II; NAD-dependent 11-beta-hydroxysteroid dehydrogenase]

[HSD11B2; HSD11B2; AME; AME1; HSD2; HSD11K; SDR9C3; HSD11K; 11-DH2; 11-beta-HSD2; -HSD11 type II; 11-beta-HSD]

DHI2_HUMAN

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

Highly purified

Supplied as a liquid in PBS, pH 7.5

1mg/ml

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.

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.

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)

Molecular Biology; MB-Enzymes, Dehydrogenase

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.

22478066

AAH36780.1

P80365

< 3kD

Aldosterone-regulated Sodium Reabsorption Pathway (130626); Aldosterone-regulated Sodium Reabsorption Pathway (130590); C21-Steroid Hormone Biosynthesis, Progesterone = Cortisol/cortisone Pathway (413395); C21-Steroid Hormone Biosynthesis, Progesterone = Cortisol/cortisone Pathway (468370); Glucocorticoid Mineralcorticoid Metabolism Pathway (198902); Prostaglandin Synthesis And Regulation Pathway (198912); Steroid Hormone Biosynthesis Pathway (82940); Steroid Hormone Biosynthesis Pathway (301)

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]

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

0.1 mg

395 USD