Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Hamster, Rabbit

Cited:

Human, Mouse, Rat, Hamster

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Proximity Ligation Assay, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all DIO3 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to an internal portion of rat DIO3 (within residues 250-300). [Swiss-Prot# P49897]

Reactivity Notes

Rabbit reactivity via customer review.

Localization

Cell membrane; Single-pass type II membrane protein

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for DIO3 Antibody - BSA Free

Western Blot: DIO3 AntibodyBSA Free [NBP1-05767]

Western Blot: DIO3 AntibodyBSA Free [NBP1-05767]

Western Blot: DIO3 Antibody [NBP1-05767] - Detection of Dio3 in rat placenta.
Flow Cytometry: DIO3 Antibody - BSA Free [NBP1-05767]

Flow Cytometry: DIO3 Antibody - BSA Free [NBP1-05767]

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 AntibodyBSA Free [NBP1-05767]

Western Blot: DIO3 AntibodyBSA Free [NBP1-05767]

Western Blot: DIO3 Antibody [NBP1-05767] - Detection of Dio3 in rabbit brain whole cell homogenate. Photo courtesy of product review by verified customer.
DIO3 Antibody - BSA Free

Western Blot: DIO3 Antibody - BSA Free [NBP1-05767] -

The effect of fibrotic-ECM and tetrac on DF cell proliferation and death and on the levels of MF biomarkers, alpha v beta 3, miRNA-2,1 and D3: DF (100,000 cells/well) were cultured on normal (−TGF beta (TGFb in the graphs)) or fibrotic-ECM (+TGF beta ) with/without 0.5 μM tetrac or its solvent (DMSO-KOH propylene glycol) for 48 h. Then, cells were harvested, and the following was undertaken: (1) the cells were counted using a counter and trypan-blue; (2) the proteins were extracted from the cells, and protein levels were measured by Western blot; and (3) the RNA was extracted from the cells, and the level of miRNA-21 was evaluated using qPCR. (A,H) shows the average number of total cells, live cells, viability, and cyclin-D1 at the end of the experiment (mean + SD). (B–F,I,J) show collagen-I (B), elastin (C), alpha SMA (D), alpha v (E), beta 3 (F), miRNA-21 (I), and D3 (J) levels, in cells cultured on ECMs with/without tetrac. (G) shows a schematic demonstration of the alpha v beta 3 binding site of RGD, T3, T4 and tetrac. (K) shows representative Western blot images of protein levels in the different treatments. The blots contain skipping lanes. * The results are significantly different (p ≤ 0.05, n = 4–6). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37240272), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
DIO3 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: DIO3 Antibody - BSA Free [NBP1-05767] -

Type 3 deiodinase expression and colocalization on muscle. (A) DIO3 expression is augmented in the muscle of the critically ill, specifically in those with NTIS. (B) panel. RNAscope showing the colocalization of DIO3 and MYOD+, augmented in patients with T3 levels < 35 ng/dL. (C) RNAscope showing that DIO3 and PAX7+ do not colocalize in patients with T3 levels < 35 ng/dL. (D) RNAscope of DIO3 and DIO2, also shows that these genes do not colocalize in patients with T3 levels < 35 ng/dL. DAPI (blue), DIO3 (green), MYOD, PAX7, DIO2. (red) * p = 0.001, ** p < 0.0001. ns: Statistically not significant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40141055), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
DIO3 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: DIO3 Antibody - BSA Free [NBP1-05767] -

Type 3 deiodinase expression and colocalization on muscle. (A) DIO3 expression is augmented in the muscle of the critically ill, specifically in those with NTIS. (B) panel. RNAscope showing the colocalization of DIO3 and MYOD+, augmented in patients with T3 levels < 35 ng/dL. (C) RNAscope showing that DIO3 and PAX7+ do not colocalize in patients with T3 levels < 35 ng/dL. (D) RNAscope of DIO3 and DIO2, also shows that these genes do not colocalize in patients with T3 levels < 35 ng/dL. DAPI (blue), DIO3 (green), MYOD, PAX7, DIO2. (red) * p = 0.001, ** p < 0.0001. ns: Statistically not significant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40141055), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for DIO3 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

reported by customer review using biotinylated antibody

Immunocytochemistry/ Immunofluorescence

reported in scientific literature (PMID 24885572)

Immunohistochemistry

reported in scientific literature (PMID 24174657)

Immunohistochemistry-Frozen

reported in scientific literature (PMID 24885572)

Immunohistochemistry-Paraffin

reported in scientific literature (PMID 25004090)

Western Blot

0.5 ug/ml
Application Notes
In Western blot, a band is seen ~31 kDa. 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.

Reviewed Applications

Read 2 reviews rated 3.5 using NBP1-05767 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: DIO3

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.

Alternate Names

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

Gene Symbol

DIO3

Additional DIO3 Products

Product Documents for DIO3 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for DIO3 Antibody - BSA Free

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.

Citations for DIO3 Antibody - BSA Free

Customer Reviews for DIO3 Antibody - BSA Free (2)

3.5 out of 5
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Customer Images

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  • Name: Linnea Vose
    Application: Western Blot
    Sample Tested: brain whole cell homogenate
    Species: Other
    Verified Customer | Posted 01/28/2013
    DIO3 Antibody - BSA Free NBP1-05767
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293 cells transfected with human D3 expression vector
    Species: Human
    Verified Customer | Posted 01/05/2010

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Protocols

View specific protocols for DIO3 Antibody - BSA Free (NBP1-05767):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for DIO3 Antibody - BSA Free

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  • Q: 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?

    A: 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.

  • Q: 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.

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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.

    A: 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.

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