TTF-1 / Thyroid-Specific TF (AA 195-245) antibody

Details for Product No. ABIN728713, Supplier: Log in to see
Antigen
Epitope
AA 195-245
7
1
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
22
10
9
1
1
1
1
Host
Rabbit
12
10
Clonality
Polyclonal
Conjugate
Un-conjugated
1
1
1
1
1
1
1
Application
Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
16
7
5
5
4
2
1
Options
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'Independent Validation' Badge
Lot Number 120319
Method validated Western Blotting
Positive Control Brain
Negative Control Liver
Notes A strong band was observed at the expected size in the positive control lysate but not in the negative control lysate.
Primary Antibody
  • Antibody: NK2 Homeobox 1 (NKX2-1) antibody
  • Catalog number: ABIN728713
  • Lot number: 120319
Secondary Antibody
  • Antibody: Goat anti-Rabbit IgG Antibody (HRP)
  • Catalog number: Log in to see
  • Lot number: YYDW62W
Controls
  • Mouse brain and liver tissue extracts were prepared using N-PER (87792 Thermo Scientific) and T-PER (78510 Thermo Scientific) protein extraction reagents, respectively.
  • Loading control: blots were stripped and re-probed for Beta-actin to ensure equal loading of lysates.
Protocol
  • 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% Beta-mercaptoethanol at 95°C for 5 minutes prior to loading.
  • 2. 24 µg of boiled extracts were loaded and resolved on a 8-16% SDS-polyacrylamide gel.
  • 3. The Spectra Multicolor Broad Range molecular mass marker (26634 Thermo Scientific) was used as a standard.
  • 4. Proteins were transferred onto PVDF membrane by tank transfer and protein transfer was confirmed with Ponceau S staining.
  • 5. The immunoblot membrane was blocked in PBS containing 3% (W/V) non-fat dry milk at room temperature for 1 hour.
  • 6. The membrane was rinsed with PBS containing 0.05% Tween-20 once.
  • 7. The membrane was immersed with the protein side up in the antibody solution in PBS containing 1% (W/V) non-fat dry milk and incubated for 2 hours at room temperature (~26°C).
  • 8. The membrane was rinsed in PBS containing 0.05% Tween-20 thrice for 10 min each.
  • 9. The membrane was incubated in the HRP-conjugated secondary antibody solution in PBS containing 1% (W/V) non-fat dry milk and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
  • 10. The membrane was rinsed in PBS containing 0.05% Tween-20 thrice for 10 min each.
  • 11. The membrane was washed in PBS twice for 30 seconds each.
  • 12. Signals were detected with Pierce ECL Western Blotting Substrate (32109, Thermo Scientific). The blot was scanned for 300 seconds.
  • 13. The membrane was rinsed three times with PBS containing 0.05% Tween-20.
  • 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
  • 15. The membrane was washed in PBS containing 0.05% Tween-20 times for 10 min each.
  • 16. Repeated Steps 5-12 with the loading control antibody (for Beta-actin) and its matching secondary antibody.
Experimental Notes None
Validation Images
Western Blotting validation image for anti-TTF-1 / Thyroid-Specific TF (AA 195-245) antibody (ABIN728713) Figure 1: Western blot analysis of mouse brain and liver extracts using NK2 Homeobox ...
Immunogen KLH conjugated synthetic peptide derived from human TTF-1
Isotype IgG
Purification Purified by Protein A.
Background

Transcription factor that binds and activates the promoter of thyroid specific genes such as thyroglobulin, thyroperoxidase, and thyrotropin receptor. Crucial in the maintenance of the thyroid differentiation phenotype. May play a role in lung development and surfactant homeostasis. Activates the transcription of GNRHR and plays a role in enhancing the circadian oscillation of its gene expression. Represses the transcription of the circadian transcriptional repressor NR1D1 (By similarity).

Subcellular location: Nucleus

Synonyms: BCH, BHC, NK-2, TEBP, TTF1, NKX2A, T/EBP, TITF1, TTF-1, NKX2.1, Homeobox protein Nkx-2.1, Homeobox protein NK-2 homolog A, Thyroid nuclear factor 1, Thyroid transcription factor 1, Thyroid-specific enhancer-binding protein, NKX2-1

Gene ID 7080
UniProt P43699
Research Area Chromatin and Nuclear Signaling, Transcription Factors
Application Notes WB 1:100-1000
FCM 1:20-100
IHC-P 1:100-500
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage -20 °C
Storage Comment Store at -20°C
Expiry Date 12 months
Supplier Images
SDS-PAGE (SDS) image for anti-TTF-1 / Thyroid-Specific TF (AA 195-245) antibody (ABIN728713) Independently Validated Antibody, image provided by Science Direct, badge number 0287...
Immunofluorescence (Cultured Cells) (IF (cc)) image for anti-TTF-1 / Thyroid-Specific TF (AA 195-245) antibody (ABIN728713) PFA fixed A549 cells with Anti-TTF1 Polyclonal Antibody at 1:200 dilution, followed ...
Flow Cytometry (FACS) image for anti-TTF-1 / Thyroid-Specific TF (AA 195-245) antibody (ABIN728713) Human A549 probed with TTF1 Polyclonal Antibody, Unconjugated (green) at 1:100 for 3...
Product cited in: Vadasz, Jensen, Moncada, Girard, Zhang, Blanchette, Finck: "Second and third trimester amniotic fluid mesenchymal stem cells can repopulate a de-cellularized lung scaffold and express lung markers." in: Journal of pediatric surgery, Vol. 49, Issue 11, pp. 1554-63, 2014 (PubMed).

Huang, Li, Lin, Shi, Lin, Li, Xu: "Upregulation of thyroid transcription factor-1 and human leukocyte antigen class I in Hashimoto's disease providing a clinical evidence for possible triggering autoimmune reaction." in: European journal of endocrinology / European Federation of Endocrine Societies, Vol. 164, Issue 5, pp. 795-800, 2011 (PubMed).