DIO3 Antibody

Images

 
Western Blot: DIO3 Antibody [NBP1-05767] - Detection of Dio3 in rat placenta.
Flow Cytometry: DIO3 Antibody [NBP1-05767] - Analysis using the Biotin conjugate of NBP1-05767. Dio3 staining of mouse hepatic stellate cell cultured for 7 days. Image from verified customer review.
Western Blot: DIO3 Antibody [NBP1-05767] - Detection of Dio3 in rabbit brain whole cell homogenate. Photo courtesy of product review by verified customer.

Product Details

Summary
Reactivity Hu, Mu, Rt, Ha, RbSpecies Glossary
Applications WB, Flow, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

DIO3 Antibody Summary

Immunogen
Synthetic peptide made to an internal portion of rat DIO3 (within residues 250-300). [Swiss-Prot# P49897]
Localization
Cell membrane; Single-pass type II membrane protein
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
DIO3
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 0.5 ug/ml
  • Flow Cytometry
  • Immunohistochemistry-Paraffin
Application Notes
This DIO3 antibody is useful for Western blot, where a band is seen ~31 kDa. Flow data from customer review using biotinylated antibody. Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 25004090) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
31 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Positive Control
Placenta Lysate (NB820-59714)
Control Peptide
DIO3 Peptide (NBP1-05767PEP)
Reviewed Applications
Read 2 Reviews rated 3.5
using
NBP1-05767 in the following applications:

Publications
Read Publications using
NBP1-05767 in the following applications:

Reactivity Notes

Rabbit reactivity via customer review.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS and 30% Glycerol
Preservative
0.1% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Alternate Names for DIO3 Antibody

  • 5DIII
  • D3
  • deiodinase, iodothyronine, type III
  • DIOIII
  • EC 1.97.1
  • EC 1.97.1.11
  • ITDI3
  • placental type
  • thyroxine deiodinase type III (selenoprotein)
  • Type 3 DI
  • type 3 iodothyronine selenodeiodinase
  • type III iodothyronine deiodinase
  • type-III 5' deiodinase
  • Type-III 5'-deiodinase

Background

DIO3 (type III iodothyronine deiodinase), an iodothyronine deiodinase family member, is expressed in placenta and several fetal tissues in the plasma/endosomal membranes as single-pass type II membrane protein, and it catalyzes the deiodination of T4 (3,5,3',5'-tetraiodothyronine) into RT3 (3,3',5'-triiodothyronine) and of T3 (3,5,3'-triiodothyronine) into T2 (3,3'-diiodothyronine), both RT3 and T2 being inactive metabolites thus preventing premature exposure of developing fetus to adult levels of thyroid hormones. DIO3 controls circulating thyroid hormone levels throughout gestation by controlling thyroid hormone inactivation during embryological development. DIO3 is homodimeric and may undergo minor heretodimerization with DIO1 or DIO2, homo- or heterodimer state being critical for its catalytic activity. Its activity has been identified in only a limited number of postnatal tissues, i.e. brain, skin, and pregnant uterus, whereas it is abundantly present in several fetal tissues. Furthermore, DIO3 re-expression has been shown in certain pathologies that include cancer, cardiac hypertrophy, myocardial infarction, chronic inflammation, and critical illness.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for DIO3 Antibody (NBP1-05767)(8)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Hamster.

We have publications tested in 5 applications: ICC/IF, IHC, IHC-Fr, IHC-P, WB.


Filter By Application
ICC/IF
(2)
IHC
(1)
IHC-Fr
(1)
IHC-P
(2)
WB
(2)
All Applications
Filter By Species
Human
(4)
Mouse
(2)
Rat
(1)
Hamster
(1)
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Showing Publications 1 - 8 of 8.
Publications using NBP1-05767 Applications Species
De Andrade PB, Neff LA, Strosova MK et al. Caloric restriction induces energy-sparing alterations in skeletal muscle contraction, fiber composition and local thyroid hormone metabolism that persist during catch-up fat upon refeeding. Front Physiol 2015 [PMID: 26441673]
Castroneves LA, Jugo RH, Maynard MA et al. Mice with hepatocyte-specific deficiency of type 3 deiodinase have intact liver regeneration and accelerated recovery from nonthyroidal illness after toxin-induced hepatonecrosis. Endocrinology 2014 Jul 08 [PMID: 25004090] (IHC-P, Mouse)

Details:
DIO3 antibody used at 1:2000 dilution for IHC-P alongwith isotype negative controls on wild-type D3 /D3flox allele and LD3KO/recombined D3null allele mice treated with CCl4. DIO3 positivity was seen in hepatocytes of D3-flox but not in LD3KO mice (Figure 2D).
IHC-P Mouse
Medina MC, Foseca T, Molina J et al. Maternal Inheritance of an Inactive Type III Deiodinase Gene Allele Affects Mouse Pancreatic b-cells and Disrupts Glucose Homeostasis Endocrinology. 2014 Jun 02 [PMID: 24885572] (IHC-Fr, ICC/IF, Mouse)

Details:
Antibody used for IHC-Fr on wild type (WT), inherited paternally/maternally (Htz-P/Htz-P) or D3 knockout (mouse with the disruption of Dio3 gene/D3KO) mouse pancreata - fixed in 4% PFA ON 4C, cryoprotected in 30% sucrose ON 4C, 8 um sections cut and immunostained with primary followed by Alexa conjugated secondary (Figure 2A and Figure 3A). D3 control peptide (NBP1-05767PEP) used as a negative control and no staining observed in antibody preincubated with the peptide. Antibody also used for ICC-IF in MIN6-C3 cells (Supplemental Figure 3)
IHC-Fr, ICC/IF Mouse
Maynard MA, Marino-Enriquez A, Fletcher JA et al. Thyroid hormone inactivation in gastrointestinal stromal tumors. N Engl J Med 2014 Apr 03 [PMID: 24693892] (IHC-P, Human)

Details:
Using the Biotin conjugated version of NBP1-05767, catalog number NBP1-05767B.
IHC-P Human
Vose LR, Vinukonda G, Jo S et al. Treatment with thyroxine restores myelination and clinical recovery after intraventricular hemorrhage. J Neurosci. 2013 Oct 30 [PMID: 24174657] (IHC, ICC/IF, Human) IHC, ICC/IF Human
Jo S, Kallo I, Bardoczi Z, Arrojo E et al. Neuronal Hypoxia Induces Hsp40-Mediated Nuclear Import of Type 3 Deiodinase As an Adaptive Mechanism to Reduce Cellular Metabolism J Neurosci 2012 Jun [PMID: 22723689] (WB, Human, Hamster) WB Human, Hamster
Kallo I, Mohacsik P, Vida B et al. A Novel Pathway Regulates Thyroid Hormone Availability in Rat and Human Hypothalamic Neurosecretory Neurons PLoS One 2012 [PMID: 22719854] (WB, Rat, Human) WB Rat, Human
Freitas BC, Gereben B, Castillo M et al. Paracrine signaling by glial cell-derived triiodothyronine activates neuronal gene expression in the rodent brain and human cells. J Clin Invest 2010 Jun [PMID: 20458138]

Details:
Using the Biotin conjugated version of NBP1-05767, catalog number NBP1-05767B.

Reviews for DIO3 Antibody (NBP1-05767) (2) 3.52

Average Rating: 3.5
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Human, Other.
We have 2 reviews tested in 1 application: WB.

Reviews using NBP1-05767:
Filter by Applications
WB
(2)
All Applications
Filter by Species
Human
(1)
Other
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot DIO3 NBP1-05767
Enlarge
reviewed by:
Linnea Vose
WB Other 01/28/2013
View

Summary

ApplicationWestern Blot
Sample Testedbrain whole cell homogenate
SpeciesOther
LotE-1
CommentsWe often get a nonspecific band ~50kD with our rabbit samples. There appears to be three rabbit bands approx. the size of Dio3, but the bottom band is brightest, and corresponds with the positive control.

Blocking

Blocking Details5% BSA in TBST, 1h 37C

Primary Anitbody

Dilution Ratio1:2000 in 2.5% BSA in TBST, overnight at 4C

Secondary Antibody

Secondary DescriptionJackson Goat anti Rabbit HRP

Details

Detection NotesECL, radiography film, 15s exposure

Comments

CommentsWe often get a nonspecific band ~50kD with our rabbit samples. There appears to be three rabbit bands approx. the size of Dio3, but the bottom band is brightest, and corresponds with the positive control.
 
reviewed by:
Anonymous
WB Human 01/05/2010
View

Summary

ApplicationWestern Blot
Sample TestedHEK293 cells transfected with human D3 expression vector
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 1% Roche Blocking solution, Blocking Time: overnight, Blocking Temp: 4 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1/2000, Incubation Dilution Buffer: 0.5% Roche blocking, Incubation Time: 2 hr, Incubation Temp: 25C

Secondary Antibody

Secondary DescriptionSecondary Ab Host Species: Roche anti-rabbit, Secondary Ab Dilution Ratio: 1/5000

Details

Detection NotesPierce, 1min Exposure, Pos Ctrl:transfected cells, Neg Ctrl:untransfected cells, Bands: Specific=~30kDa, Non-Specific=many

Product General Protocols

View specific protocols for DIO3 Antibody (NBP1-05767): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for DIO3 Antibody (NBP1-05767). (Showing 1 - 2 of 2 FAQs).

  1. I am looking for a Dio3 peptide that I can inject in vivo to produce a biological effect. Can you provide information or confirm whether the Dio3 peptide you have would be effective as a biological agent in vivo?
    • I have pulled up information on the Dio3 peptides and recombinant proteins that we have available. Unfortunately I would not expect any of them to be biologically active and capable of producing a response when injected in vivo. The two peptides we carry are simply intended to be used for negative control competition assays to block signal of the primary and determine what is specific and non-specific as far as signal is concerned. The other two recombinant proteins are ones we distribute for a Taiwanese company called Abnova and they should not be biologically active based on the cell free wheat germ system employed to synthesize them. We will typically list our active proteins with an indication of such on our datasheet if it has been tested and usually the application of functional listed.
  2. We have used one of your antibodies against Dio3 (cat# NBP1-05767) in the past and, despite it being directed against rodent D3 peptide sequence, it also appears to stains human D3 specifically in our hands. Can you please let us know the rodent D3 peptide sequence that was used to create this antibody? Alternatively, can you let us know if this rodent D3 peptide sequence also matches the human D3 peptide sequence? If there is a high degree of sequence homology, we wish to publish our findings and include a description that your antibody works in human tissues (IHC).We also noticed a new D3 aB on your website (cat# NBP1-19747) that is said to recognize human D3. Can you also let me know the sequence of the peptide used to generate it? We are one of the only labs in the world that study D3. If you can send us some samples of these antibodies, we can likely help you validate them experimentally for Western, IHC, etc.
    • The immunogen shares 93% homology to human. It is expected to cross-react with human cells. Furthermore, full-length DIO3 in human and mouse share greater than 95% similarity over the entire protein. It will be very difficult to find an antibody that works on one species but not the other unless the immunogen falls within amino acids 48-55, which is the only region where there is significant difference between the two species. It is highly doubtful you will find a commercial antibody that uses this immunogen to generate their antibody as most companies will want to increase the amount of species recognized by an antibody, not decrease it. For NBP1-19747, like all of our immunogens, the exact sequence is considered proprietary, but as you can see on our datasheet, the immunogen surrounds amino acid 66, which does not fall within the range where mouse and human are dissimilar.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NBP1-05767AF405
Alexa Fluor 488 Labeled NBP1-05767AF488
Alexa Fluor 647 Labeled NBP1-05767AF647
Alexa Fluor 700 Labeled NBP1-05767AF700
Biotin Labeled NBP1-05767B
DyLight 405 Labeled NBP1-05767V
DyLight 488 Labeled NBP1-05767G
DyLight 550 Labeled NBP1-05767R
DyLight 650 Labeled NBP1-05767C
FITC Labeled NBP1-05767F
HRP Labeled NBP1-05767H

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Recent Reviews

3.5
5
0
4
1
3
1
2
0
1
0

Linnea Vose
01/28/2013
Application: WB
Species: Other

 
Anonymous
01/05/2010
Application: WB
Species: Human

Bioinformatics

Gene Symbol DIO3