Overview

  • Product name
  • Description
    Mouse monoclonal to HSD3B1
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human HSD3B1 aa 1-373.

Properties

Applications

Our Abpromise guarantee covers the use of ab55268 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-P Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration. PubMed: 21094146

Target

  • Function
    3-beta-HSD is a bifunctional enzyme, that catalyzes the oxidative conversion of Delta(5)-ene-3-beta-hydroxy steroid, and the oxidative conversion of ketosteroids. The 3-beta-HSD enzymatic system plays a crucial role in the biosynthesis of all classes of hormonal steroids. Efficiently catalyzes the transformation of pregnenolone to progesterone, 17-alpha-hydroxypregnenolone to 17-alpha-hydroxyprogesterone, DHEA to 4-androstenedione, dihydrotestosterone to 5-alpha-androstane-3 beta,17 beta-diol, dehydroepiandrosterone to androstenedione and 5-alpha-androstan-3 beta,17 beta-diol to 5-alpha-dihydrotestosterone.
  • Tissue specificity
    Placenta and skin. Predominantly expressed in mammary gland tissue.
  • Pathway
    Lipid metabolism; steroid biosynthesis.
  • Sequence similarities
    Belongs to the 3-beta-HSD family.
  • Cellular localization
    Endoplasmic reticulum membrane. Mitochondrion membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 3 beta and steroid delta isomerase 1 antibody
    • 3 beta and steroid hydroxy delta 5 steroid dehydrogenase antibody
    • 3 beta HSD I antibody
    • 3 beta-hydroxysteroid dehydrogenase/Delta 5-->4-isomerase type I antibody
    • 3-beta-HSD I antibody
    • 3-beta-hydroxy-5-ene steroid dehydrogenase antibody
    • 3BETAHSD antibody
    • 3BH antibody
    • 3BHS1_HUMAN antibody
    • 3BHSD antibody
    • Delta-5-3-ketosteroid isomerase antibody
    • HGNC:5217 antibody
    • HSD3B antibody
    • HSD3B1 antibody
    • HSDB3 antibody
    • HSDB3A antibody
    • Hydroxy delta 5 steroid dehydrogenase 3 beta and steroid delta isomerase 1 antibody
    • Hydroxy delta 5 steroid dehydrogenase antibody
    • Progesterone reductase antibody
    • SDR11E1 antibody
    • Steroid Delta-isomerase antibody
    • trophoblast antigen antibody
    • Trophoblast antigen FDO161G antibody
    see all

Images

  • HSD3B1 antibody (ab55268) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human placenta.
  • Anti-HSD3B1 antibody (ab55268) at 1 µg/ml + Human placenta tissue lysate - total protein (ab29745) at 10 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa
    Additional bands at: 73 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes
  • Overlay histogram showing JEG3 cells stained with ab55268 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55268, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in JEG3 cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Badr G  et al. Protective effects of camel whey protein against scrotal heat-mediated damage and infertility in the mouse testis through YAP/Nrf2 and PPAR-gamma signaling pathways. Mol Reprod Dev 85:505-518 (2018). Read more (PubMed: 29683243) »
  • Xie T  et al. Roscovitine protects murine Leydig cells from lipopolysaccharide-induced inflammation. Exp Ther Med 13:2169-2176 (2017). Read more (PubMed: 28565824) »
See all 9 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Pig Tissue sections (Ovary)
Permeabilization
No
Specification
Ovary
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Ms. Natasja Costermans

Verified customer

Submitted Nov 09 2018

Question
Answer

Anti-HSD3B1 antibody (ab55268) is raised against the full length protein, corresponding to amino acids 1-374 of Human HSD3B1.

Sequence alignment of Human HSD3B1 against HSD3B2 is 94%.

This suggests that ab55268 is not specific to HSD3B1 and will cross react with HSD3B2.  As the sequence homology is so similar, it may difficult to find an antibody specific to HSD3B1.

Read More

Answer

Thank you for your reply.

We recommend using 5% milk or BSA for the blocking step, however, when incubating with the primary antibody we recommend reducing that concentration to 1% or less. Blocking agent is not strictly necessary in the antibody diluent and can reduce the signal when used at higher concentrations.

I hope that this will help to improve our customer's results. If not, please let me know and I will be happy to offer either a free of charge replacement or a credit.

Read More

Answer

Thank you for contacting us and for forwarding the customer's protocol information. I am sorry that this antibody is not producing satisfactory results in WB. Could you please provide some additional information about the experiment?

1) Were the samples used human? Could you please let me know what samples are present in each lane of the first image using ab55268? It appears that there may be a band in the last lane.

2) When the customer reports using BSA and milk at 5%, was that 5% BSA and 5% milk in the same blocking buffer solution, or did the customer try each separately? Was the percentage also 5% for the antibody diluent? For the antibody diluent I typically recommend a blocking agent concentration less than 1%. High concentrations of other proteins can interfere with antibody binding and cause a loss of signal.

3) Could you please confirm the PO number or order reference number, or let me know what date the antibody was purchased? I have been unable to locate any orders of this antibody from your company.

I hope this will help to improve the customer's results. If not, please let me know and I will be happy to help you further.

Read More

Answer

Thank you for filling in the questionnaire and for reporting this problem to us. We will use the details you sent about your protocol in order to investigate the source of the problem. I have looked through the protocol, which looks fine to me. I appreciate the time that has been spent on these experiments and would be pleased to arrange a a credit note or a replacement with the alternative Mouse monoclonal HSD3B1 antibody ab69807 that has been tested in IHC-P and Human. The link to the datasheet of ab69807 is www.abcam.com/ab69807 I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for contacting us. The HSD3B1 protein has 71% similarity between human and mouse. Theoretically the antibody should not be detecting the mouse protein however because we have not tested this so will not be able to confirm it. http://www.ebi.ac.uk/Tools/services/web/toolresult.ebi?jobId=clustalw2-I20110823-120040-0090-47470728-pg&tool=clustalw2&analysis=alignments If customer is happy using this ab with mouse lysates please let me know, I will be happy to send them an Abreview testing discount code. I would recommend using human lysates as positive control along with mouse lysates. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Testis)
Specification
Testis
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 09 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Placenta)
Specification
Placenta
Fixative
Formaldehyde
Antigen retrieval step
None
Permeabilization
No

Abcam user community

Verified customer

Submitted Jul 20 2009

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

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