- Tested Species Reactivity:
- Human, Monkey
- Predicted Species Reactivity:
- Goat, Human, Monkey
- Product Format:
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
- Reconstitution and Storage:
- For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
- Replacement Item:
- This antibody may replace item sc-100466 from Santa Cruz Biotechnology.
- The immunogen is a synthetic peptide directed towards the N terminal region of human HSD3B1
- Affinity Purified
- Predicted Homology Based on Immunogen Sequence:
- Goat: 82%; Human: 100%; Monkey: 92%
- Complete computational species homology data:
- Anti-HSD3B1 (ARP41821_P050)
- Peptide Sequence:
- Synthetic peptide located within the following region: TGWSCLVTGAGGFLGQRIIRLLVKEKELKEIRVLDKAFGPELREEFSKLQ
- Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
- Blocking Peptide:
- For anti-HSD3B1 (ARP41821_P050) antibody is Catalog # AAP41821 (Previous Catalog # AAPP10869)
- Printable datasheet for anti-HSD3B1 (ARP41821_P050) antibody
- Target Reference:
- Ross,R.W., (2008) J. Clin. Oncol. 26 (6), 842-847
Mirkheshti, N; Park, S; Jiang, S; Cropper, J; Werner, SL; Song, CS; Chatterjee, B; Dual targeting of androgen receptor and mTORC1 by salinomycin in prostate cancer. 7, 62240-62254 (2016). WB, Cow, Goat, Horse, Human, Pig, Rat, Sheep, Monkey 27557496
- Gene Symbol:
- Official Gene Full Name:
- Hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1
- Alias Symbols:
- HSD3B, HSDB3, I, HSDB3A, SDR11E1, 3BETAHSD
- NCBI Gene Id:
- Protein Name:
- 3 beta-hydroxysteroid dehydrogenase/Delta 5-->4-isomerase type 1
- Description of Target:
- 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.
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Protein Size (# AA):
- Molecular Weight:
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express HSD3B1.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express HSD3B1.
We used the anti-HSD3beta-1 antibody, rabbit polyclonal from Aviva System Biological to probe 3-beta HSD1 in lysates of human prostate cancer cell lines. Compared to other vendors, this antibody from Aviva works better. The antibody does generate non-specific bands along with the 42 kDa band for 3-beta HSD1. Conditions for gel run and membrane washing can be standardized to improve the quality of Western blot data. We published an article in 2016 containing data that utilized this antibody.
Researcher: Jonathan Bertin, Endoceutics Inc.
Application: Western Blotting
Species + Tissue/Cell type: Lane 1: 50ug monkey brain extract
Primary antibody dilution: 1:1000
Secondary antibody: Goat anti rabbit-HRP
Secondary antibody dilution: 1:10,000
|How do Aviva's reagents play a role in your experimental goals?||The antibody was utilized to detect the protein in monkey brain tissue. Giving the high homology between monkey and human sequences of the protein, we were able to specifically detect a low level of the targeted enzyme by western blot. We are now confident to utilize the ARP41821 antibody for immunohistrochemistry work to detect the spatial distribution of the enzyme in monkey brain tissue.|
|How would you rate this antibody on a scale from 1-5 (5=best) nad why?||5|
|Would you use this antibody in future experiment?||Absolutely. It will now be tested for IHC work once again in monkey brain tissue.|
|Have you used another antibody which has worked in your application?||Yes. Homemade rabbit polyclonal antibody which as revealed to be somewhat unspecific.|
|Do you believe the information about the reagent on Aviva's website is correct?||Yes|
|If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?||If it now works in IHC in monkey brain tissue (paraffin), we will undoubtedly publish the results.|
|How did you store the antibody after re-suspension?||After resuspension at 0.2 mg/mL, it was kept at 4 degree Celsius.|
|Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):||Normal monkey (cynomolgus) brain tissue. 50ug of total protein were loaded per well.|
|How many different experimental trials were conducted using the antibody sample?||Only one.|
|How was this sample prepared?||Total proteins were quantified and the sample was diluted in Laemmli buffer. 50ug of proteins were migrated on a SDS-PAGE gel.|
|Primary antibody dilution and incubation time:||1/200 in 2.5% milk buffer. Incubation overnight at 4 degree Celsius.|
|Secondary antibody used and dilution and incubation time:||HRP-conjugated goat anti-rabbit. 1/10000 in 2.5% milk buffer. Incubation room temperature 1 hour.|
|What controls were used in your experiment (positive/negative)?||Negative. Membranes blotted using only secondary antibody. Revealed no unspecific staining by the secondary antibody.|
|Please include your detailed WB Procedure/Protocol here:||A mid-brain coronal cut was done with a snap frozen bloc of cynomolgus brain tissue utilizing a cryostat. The slice weigh
was 100ug. Total proteins were extracted with lysis buffer (6M urea, 20 mM Tris pH 6.8 and 1% SDS). After protein
quantification, samples were diluted in 2X laemmli buffer. 50ug of proteins were migrated on a 10% acrylamide SDS-PAGE gel (70V for 3 hours) and then transferred on a PVDF membrane (100V for 1 hour). The membrane was then incubated with the primary and HRP-secondary antibodies as described previously before being incubated for 1 min with Bio-Rad Clarity™ Western ECL Substrate before being revealed on film (10 min exposition). Membrane was washed 3 times/10 min each in 0.05% NP40-Tween20 buffer after primary and secondary antibody incubations.