TTF1 antibody (AA 201-300)

Catalog No. ABIN728713

This independently validated Rabbit Polyclonal antibody specifically detects TTF1 in WB, FACS, ICC and IF (cc). It exhibits reactivity toward Human, Rat and Mouse and has been mentioned in 2+ publications.

Quick Overview for TTF1 antibody (AA 201-300) (ABIN728713)

Target: TTF1 (Transcription Termination Factor, RNA Polymerase I (TTF1))

Reactivity: Human, Rat, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This TTF1 antibody is un-conjugated

Application: Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (Cultured Cells) (IF (cc))

Product Details anti-TTF1 Antibody

Binding Specificity: AA 201-300

Cross-Reactivity: Human, Mouse, Rat

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human TTF-1

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (WB)

Validation #028752 (Western Blotting)

by: Alamo Laboratories Inc

No.: #028752

Date: 09/08/2013

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.

Validation Images

Figure 1: Western blot analysis of mouse brain and liver extracts using NK2 Homeobox 1 (NKX2-1) antibody (Catalog number ABIN728713, Lot number 120319). NKX2-1 is present in the positive control sample (brain) and absent from the negative control sample (liver). The arrowhead indicates the expected position of NKX2-1 (predicted MW ~38kDa). 24 micrograms of total protein extracts from each sample were loaded into each lane. Upper panel: scanned image of the NKX2-1 antibody probed with the liver and brain extracts in lanes 2 and 3, respectively. Lower panel: scanned image of the loading control (Beta-actin).

Full Methods

Primary Antibody:

• Antibody: NK2 Homeobox 1 (NKX2-1) antibody
• Catalog number: ABIN728713
• Lot number: 120319

Secondary Antibody:

• Antibody: Goat anti-Rabbit IgG Antibody (HRP)
• Lot number: YYDW62W

Full 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

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-TTF1 antibody (ABIN728713)

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).

Target Details for TTF1

Target: TTF1 (Transcription Termination Factor, RNA Polymerase I (TTF1))

Alternative Name: TTF1

Background:

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

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).

Gene ID: 7080

UniProt: P43699

Pathways: Thyroid Hormone Synthesis, Regulation of Systemic Arterial Blood Pressure by Hormones, Feeding Behaviour