TTF1 antibody (AA 201-300)

Catalog No. ABIN728713

Product Details anti-TTF1 Antibody

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

Binding Specificity: AA 201-300

Reactivity: Human, Rat, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This TTF1 antibody is un-conjugated

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

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 (TTF1 Products)

Synonyms: TTF-1 antibody, TTF-I antibody, AV245725 antibody, RGD1565673 antibody, transcription termination factor 1 antibody, transcription termination factor, RNA polymerase I antibody, TTF1 antibody, Ttf1 antibody

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