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Thyroglobulin

IHC of Thyroglobulin on an FFPE Thyroid Tissue

Description

Thyroglobulin (Tg) is a 660 kDa, dimeric protein produced by and used entirely within the thyroid gland. Tg is used by the thyroid gland to produce the thyroid hormones thyroxine (T4) and triiodothyronine (T3). The active form of thyroxine, triiodothyronine, is produced both within the thyroid gland and on the periphery by 5’-deiodinase, which has been referred to as Tetraiodothyronine-5-deiodinase.


This antibody reacts with human thyroglobulin as demonstrated by a single band of immunoblotting in a lysate of human thyroid tissue. The vast majority of follicular carcinomas of the thyroid will give positive immunoreactivity for thyroglobulin, sometimes only focally. Poorly-differentiated Carcinomas of the Thyroid are frequently thyroglobulin negative. Adenocarcinomas of non-thyroid origin do not react with this antibody.

Antibody Type
Mouse Monoclonal
Clone
2H11 + 6E1
Isotype
IgG1
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Thyroid
Storage
Store at 2°-8°C
Stability
2 years

For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).

Presentation

Thyroglobulin is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5973
Prediluted
Ready-To-Use
3.0 ml
BSB 5974
Prediluted
Ready-To-Use
7.0 ml
BSB 5975
Prediluted
Ready-To-Use
15.0 ml
BSB 5976
Concentrated
1:100-1:500
0.1 ml
BSB 5977
Concentrated
1:100-1:500
0.5 ml
BSB 5978
Concentrated
1:100-1:500
1.0 ml
BSB 5979
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Bellet D, et al. J Clin Endocrin Metab. 1983;56:530-533

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
  5. Any of three heating methods may be used:
    • Electric Pressure Cooker
      Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
    • Water Bath Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
    • Conventional Steamer Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain