Overview

  • Product name
  • Description
    Rabbit polyclonal to HSD11B1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Horse, Human
    Predicted to work with: Rabbit, Hamster, Cow, Cat, Dog, Monkey, Cynomolgus monkey, Macaque monkey
  • Immunogen

    Synthetic peptide:

    CLELGAASAHYIAGT

    , corresponding to amino acids 78/92 of Human HSD11B1

  • Positive control
    • Recombinant Human HSD11B1 protein (ab114301) can be used as a positive control in WB. Rat and mouse liver microsomes.

Properties

Applications

Our Abpromise guarantee covers the use of ab39364 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).Can be blocked with HSD11B1 peptide (ab101097).
IHC-P Use a concentration of 2 µg/ml.
IHC-Fr Use at an assay dependent concentration. PubMed: 20385225

Target

  • Function
    Catalyzes reversibly the conversion of cortisol to the inactive metabolite cortisone. Catalyzes reversibly the conversion of 7-ketocholesterol to 7-beta-hydroxycholesterol. In intact cells, the reaction runs only in one direction, from 7-ketocholesterol to 7-beta-hydroxycholesterol.
  • Tissue specificity
    Widely expressed. Highest expression in liver.
  • Involvement in disease
    Defects in HSD11B1 are a cause of cortisone reductase deficiency (CRD) [MIM:604931]. In CRD, activation of cortisone to cortisol does not occur, resulting in adrenocorticotropin-mediated androgen excess and a phenotype resembling polycystic ovary syndrome (PCOS).
  • Sequence similarities
    Belongs to the short-chain dehydrogenases/reductases (SDR) family.
  • Post-translational
    modifications
    Glycosylated.
  • Cellular localization
    Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 11 beta HSD 1 antibody
    • 11 beta HSD1 antibody
    • 11 beta hydroxysteroid dehydrogenase 1 antibody
    • 11 DH antibody
    • 11-beta hydroxysteroid dehydrogenase, type 1 antibody
    • 11-beta-HSD1 antibody
    • 11-beta-hydroxysteroid dehydrogenase 1 antibody
    • 11-DH antibody
    • 11DH antibody
    • Corticosteroid 11 beta dehydrogenase isozyme 1 antibody
    • Corticosteroid 11-beta-dehydrogenase isozyme 1 antibody
    • CORTRD2 antibody
    • DHI1_HUMAN antibody
    • HDL antibody
    • HSD 11 antibody
    • HSD11 antibody
    • HSD11B antibody
    • HSD11B1 antibody
    • HSD11L antibody
    • Hydroxysteroid (11 beta) dehydrogenase antibody
    • Hydroxysteroid (11 beta) dehydrogenase 1 antibody
    • MGC13539 antibody
    • SDR26C1 antibody
    • short chain dehydrogenase/reductase family 26C, member 1 antibody
    see all

Images

  • Western blot analysis of Sheep liver tissue lysate (5µg/lane) labeling HSD11B1 with ab39364 at 1/500 (16 hours at 4ºC). An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Anti-HSD11B1 antibody (ab39364) at 1 µg/ml + Human liver tissue lysate - total protein (ab29889) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 32 kDa


    Exposure time: 16 minutes


    HSD11B1 contains a number of potential glycosylation sites (SwissProt) which may explain the higher migration pattern observed.

References

This product has been referenced in:
  • Ghnenis AB  et al. Maternal obesity in the ewe increases cardiac ventricular expression of glucocorticoid receptors, proinflammatory cytokines and fibrosis in adult male offspring. PLoS One 12:e0189977 (2017). Read more (PubMed: 29267325) »
  • Doig CL  et al. 11ß-HSD1 Modulates the Set Point of Brown Adipose Tissue Response to Glucocorticoids in Male Mice. Endocrinology 158:1964-1976 (2017). Read more (PubMed: 28368470) »
See all 14 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris ⁄EDTA
Sample
Horse Tissue sections (Testicular tissue)
Specification
Testicular tissue
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 30 2014

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Sample
Sheep Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Dr. Rob Cordery-Cotter

Verified customer

Submitted Nov 12 2013

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of a new lot.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

rfor kindly completing the questionnaire.

Reviewing theimage, despite the nonspecific bands there is a well resolved band at the expected molecular weight and it is odd that the blocking peptide did not complete obliterate this band as it had previously. Again, as previously discussed this is difficult to explain.
I understand your concerns, and it is regrettable the results have not been satisfactory for you. Iwould like to offer the following considerations:

1. The blocking peptide should be adjusted in terms of concentration andif youused a 5 fold excess, they may want to try a 10 fold excess which is often what we currently suggest. The blocking peptide should block only HSD1 protein.

2. Possibly there could be another peptide in the sample which is not HSD1 and happens to bind at 34 kDa and thus is still present upon use of the blocking peptide. If the blocking peptide and antibody have been stored for 2 ½ years it is possible that some degradation has occurred also, especially since the experiment was satisfactory at the onset.

3. Could you confirm what dilution of antibody has been tried? We recommend to dilute at1:200 for this application, although this may require some further optimization.

4. Could you confirm the antibody was aliquoted beforefreezing? This is important in order to avoid freeze thaw cycles which can damage the antibody.

As previously discussed, our guarantee period is 6 months. I am sorry, but at this late date after purchase we are regrettably not able to offer a replacement in this case. Wewould encourage customers to contact us as soon as possibleif they experience any difficulties, and to contact us within 6 months.

As a gesture of goodwill on this occasion, I can offer you a code to obtain 5% off your next antibody purchase, and free shipping. i hope this would be helpful to you. In order to make use of this offer, please provide the following codes with your order:

5% discount: 5PC-OFF-T7T62
Free shipping: NO_PORTO-T7T61

Thank you for your cooperation,I would be pleased and interested to hear from you with the results of the suggestions made. If you have any further questions or concerns regarding this case, please do not hesitate to contact us.

Read More

Question

Sir/Madam,

We have bougthHSD11B1 antibody (ab39364) from your Company in December 2010. We have tried this antibody on microsomes from rat liver and have obtained very interesting results: on the same samples of liver microsomes one time we got a double band on approximatelly 34 kDa and one band on 43 kDa (Picture 1, A), while other time on the same samples we obtained one band on 34 kDa and bands on app. 53 and 43 kDa (Picture 2, A).At the sametime on microsomes from rat adipose tissue we got one bend on 34 kDa (picture 3). Since we were not assure about specificity of our bands in liver samples, we bought a blocking peptide for this antibody (ab101097) from Institute for Nuclear Sciences VINCAthrough PIU tender. We prepared experiment with blocking peptide as you suggested in your protocol (BLOCKING WITH IMMUNIZING PEPTIDE (BL) PROTOCOL) using final concentration of HSD1 antibody 1 microgr/ml and blocking peptide 5 microgr/ml (5 fold excess as you suggested in peptide competition assay). In the first experiment (Picture 1, B) using of blocking peptide led to disappearance all bends, while in other experiment (Picture 2, B) specific band on 34 kDa showed faint signal while other bands totally dissapeared.

Questions:

1. Does HSD1 antibody (ab39364) recognize HSD2, since it is on 44 kDa and due to high similarity between HSD1 and HSD2?

2. Is the protocol with blocking peptide correct, should I use lesser concentration of blocking peptide (for example 2 fold excess) to obtain better results?

3. In case od double bands on 34 kDa, which band I should use as specific?

As evidence I enclosed scan images ofthree different Western blot membranes probed with this antibody.

Thank you in advance forquick answer.

Best regards,

Read More
Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and forMouse, Rat andHuman samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

In answer to your questions:

1. Does HSD1 antibody (ab39364) recognize HSD2, since it is on 44 kDa and due to high similarity between HSD1 and HSD2?

I can confirm we are not aware of any data to suggest that this antibody would show cross reactivity to recognize HSD2.

2. Is the protocol with blocking peptide correct, should I use lesser concentration of blocking peptide (for example 2 fold excess) to obtain better results?

I can suggest to consider using a higher concentration of blocking peptide as the blockingmay not becomplete.

You mention that in the first experiment use of blocking peptide led to disappearance all bands, while in other experiment at 34 kDa there is a faint signal. I am sorry I do not know how to explain this difference between two uses of the same antibody and the same blocking peptide, except that this may indicate incomplete blocking.

3. In case of double bands on 34 kDa, which band I should use as specific?

This would be difficult to answer. As the results show nonspecific bands,in order to identify those bands in the rat liver microsome samples further experiments would be necessary. Using a higher concentration of blocking peptide may help in this case so you can observe which of the bands remains.

This antibody ab39364 has been confirmed to be cross reactive with rat 11β-HSD1 and the expected band should appear at 32 kDa. Unfortunately, I also did not find anything in literature to suggest there are any different size isoforms of SD11B1 in liver samples. You may like to try a more extensive literature search to confirm this.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would also appreciate if you could confirm: if the same samples used in both experiments?

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Question
Answer

Thank you for contacting us.

Your credit note ID is ************.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up