Overview

  • Product name

  • Description

    Rabbit polyclonal to HSD17B6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Horse, Guinea pig, Cow, Cat, Dog
  • Immunogen

    Synthetic peptide corresponding to a sequence within N terminal amino acids 1-50 of Human HSD17B6 (MWLYLAAFVG LYYLLHWYRE RQVVSHLQDK YVFITGCDSG FGNLLARQLD)

  • Positive control

    • A172 cells. Liver tissue.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituents: 2% Sucrose, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab62221 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 1.25 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.

Target

  • Function

    NAD-dependent oxidoreductase with broad substrate specificity that shows both oxidative and reductive activity (in vitro). Has 17-beta-hydroxysteroid dehydrogenase activity towards various steroids (in vitro). Converts 5-alpha-androstan-3-alpha,17-beta-diol to androsterone and estradiol to estrone (in vitro). Has 3-alpha-hydroxysteroid dehydrogenase activity towards androsterone (in vitro). Has retinol dehydrogenase activity towards all-trans-retinol (in vitro). Can convert androsterone to epi-androsterone. Androsterone is first oxidized to 5-alpha-androstane-3,17-dione and then reduced to epi-andosterone. Can act on both C-19 and C-21 3-alpha-hydroxysteroids.
  • Tissue specificity

    Detected in liver and prostate (at protein level). Detected in adult liver, lung, brain, placenta, prostate, adrenal gland, testis, mammary gland, spleen, spinal cord and uterus. Detected in caudate nucleus, and at lower levels in amygdala, corpus callosum, hippocampus, substantia nigra and thalamus. Detected in fetal lung, liver and brain.
  • Sequence similarities

    Belongs to the short-chain dehydrogenases/reductases (SDR) family.
  • Cellular localization

    Microsome membrane. Early endosome membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • 17-beta-HSD 6 antibody
    • 17-beta-HSD6 antibody
    • 17-beta-hydroxysteroid dehydrogenase type 6 antibody
    • 3 hydroxysteroid epimerase antibody
    • 3(alpha >beta) hydroxysteroid epimerase antibody
    • 3(alpha >beta) hydroxysteroid epimerasel antibody
    • 3-alpha->beta-HSE antibody
    • 3-alpha->beta-hydroxysteroid epimerase antibody
    • H17B6_HUMAN antibody
    • HSD17B6 antibody
    • HSE antibody
    • Hydroxysteroid (17 beta) dehydrogenase 6 antibody
    • Hydroxysteroid (17 beta) dehydrogenase 6 homolog (mouse) antibody
    • Hydroxysteroid 17 beta dehydrogenase 6 antibody
    • NAD+ dependent 3 alpha hydroxysteroid dehydrogenase 3 hydroxysteroid epimerase antibody
    • NAD+ dependent 3 alpha hydroxysteroid dehydrogenase antibody
    • Oxidative 3 alpha hydroxysteroid dehydrogenase antibody
    • Oxidative 3-alpha hydroxysteroid dehydrogenase antibody
    • Oxidoreductase antibody
    • Retinol dehydrogenase antibody
    • RODH antibody
    • SDR9C6 antibody
    • Short chain dehydrogenase/reductase family 9C, member 6 antibody
    see all

Images

  • Anti-HSD17B6 antibody (ab62221) at 1.25 µg/ml + A172 cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 35 kDa
    Observed band size: 35 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling HSD17B6 with ab62221 at 4-8µg/ml. Arrows indicate positively labelled hemopoietic cells. Magnification: 400X.

References

This product has been referenced in:

  • Mohler JL  et al. Activation of the androgen receptor by intratumoral bioconversion of androstanediol to dihydrotestosterone in prostate cancer. Cancer Res 71:1486-96 (2011). WB . Read more (PubMed: 21303972) »
  • Muthusamy S  et al. Estrogen receptor ß and 17ß-hydroxysteroid dehydrogenase type 6, a growth regulatory pathway that is lost in prostate cancer. Proc Natl Acad Sci U S A 108:20090-4 (2011). IHC-P ; Human . Read more (PubMed: 22114194) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Question
Answer

The peptide to ab62221 has been added as HSD17B6 peptide (ab156928)- https://www.abcam.com/hsd17b6-peptide-ab156928.html

Read More

Answer

Thank you for your response and for attaching the Western blot images.

There is a clear difference in the banding pattern between cell prepared with RIPA buffer and those with Laemmli lysis buffer. This highlights the importance of use of proteinase inhibitors in the lysis buffer during sample preparation to prevent protein degradation.

On the left image (when RIPA used), there is a distinctive single band in liver tissue lysates and a double band in the PC3 cell lysate just below 50 kDa. This may well be the signal for HSD17B6. The shift between the expected band size (36 kDa) and the actual size (48 kDa) is very likely due to post-translational modification (i.e. glycosylation).

On the right image (Laemmli), in some lanes there are hardly any visible bands or bands appear at lower size 26 kDa than the expected theoretical size.

Swiss-Prot database, the predicted band size of HSD17B6 is 36 kDa. However, it is important to emphasize that there are 3 potential N-glycosylation sites at 161, 215 and 256 aa along the polypeptide chain. You can find some further information about the processed protein at this site:

http://www.uniprot.org/uniprot/O14756

I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Question

Dear Sir/Madam

Here I sent report form and the last experiment. It seems that this antibody didn’t work well. I put the picture in pdf file. Thank you. Best Regard



1) Abcam product code ab62221



2) Abcam order reference number or product batch number

Lot 858589

3) Description of the problem



Apparently, this antibody is not detected expected potein



4) Sample preparation:

Type of sample : whole cell lysate

Lysis buffer : Laemli buffer ( 1mM CaCl2, 2% SDS, 60 mM tris Glycine pH 6,8)

Protease inhibitors : No

Phosphatase inhibitors : No

Reducing agent : B- mercaptoethanol

Boiling for ≥5 min? : yes

Protein loaded : 10 ug/lane or cells/lane

Positive control : Liver

Negative control : we still figure it out



5) Percentage of gel

Type of membrane : 10 % SDS Page

Protein transfer verified : PVDF membrane

Blocking agent and concentration : 0.5% milk in TBS-0.1% Tween

Blocking time : 1 hour

Blocking temperature : room temperature



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer : TBS - 0.1% Tween

Incubation time : 24 hour

Incubation temperature : 4oC



7) Secondary antibody:

Species:

Reacts against : Rabbit

Concentration or dilution : 1: 50.000

Diluent buffer : TBS - 0.1% Tween

Incubation time : 1 hour

Incubation temperature : room temperature

Fluorochrome or enzyme conjugate: no



8) Washing after primary and secondary antibodies:

Buffer : TBS - 0.1% Tween

Number of washes : 3 times



9)Detection method

ECL kit

10) How many times have you run this staining?

4 times

Do you obtain the same results every time?

Yes

What steps have you altered to try and optimize the use of this antibody?

Increase prim Antibody concentration

Increase prim Antibody incubation time

Read More
Answer

Thank you for your enquiry regarding ab62221 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

1) Band size:

According to Swiss-Prot database, the predicted band size of HSD17B6 is 36 kDa, you can find some further information about the processed protein at this site:

http://www.uniprot.org/uniprot/O14756

2) Lysis buffer:

It may well be that the target protein is degraded during sample preparation since no proteinase inhibitors have been used the lysis buffer. RIPA buffer is recommended to use for lysing the cells/tissues which contains a cocktail of different proteinase inhibitors to prevent any unwanted degradation. The recipe of the RIPA buffer can be found under our Protocol site:

https://www.abcam.com/index.html?pageconfig=resource&rid=11379#A1

3) Optimization:

3.a Loading different amount of protein i.e. 15, 20,25 ug per lane would be worth trying to see if the signal is getting stronger or not.

3.b Testing diffrent dilutions of primary antibody i.e. 1/500 or 1/250

3.c HSD17B6 is expressed in microsomal and early endosomal membranes, so it would be worth using membrane enriched lysates - instead of whole tissue lysates.

4) Loading control:

It would be worth running a loading control to make sure that the sample preparation and the loading was sufficient.

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

Read More

Answer

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

We are expecting the return of the 2 vials of ab62221 to our Cambridge, MA office. As requested, I have issued a free of charge replacement of 2 vials of ab88892. These are backordered, but you can expect them on Feb 20th. 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

Thank you for your enquiry regarding ab62221. The expected band size of HSD17B6 protein is around 35 kDa, however several factors may influence the actual band size. Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response. I look forward to hearing from you so

Read More

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