mtTFA Recombinant Rabbit Monoclonal Antibody [JM32-44]
cat.: ET1705-64
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | JM32-44 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 29 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human mtTFA aa 41-90 / 246. |
| Positive control: | K-562 cell lysate, HL-60 cell lysate, HeLa cell lysate, HepG2 cell lysate, MCF7 cell lysate, Human brain tissue lysate, human kidney tissue, human colon tissue, HeLa, human colon cancer tissue. |
| Subcellular location: | Mitochondrion, Mitochondrion matrix, mitochondrion nucleoid. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P IP |
1:1,000 1:100-1:200 1:50-1:200 1:200-1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: Q00059 Human |
| Alternative names: | anscription factor 6-like 1 Mitochondrial transcription factor 1 mitochondrial transcription factor A MtTF1 mtTFA TCF 6 TCF-6 TCF6 TCF6L1 TCF6L2 TCF6L3 TFAM TFAM_HUMAN Transcription factor 6 Transcription factor 6 like 2 (mitochondrial transcription factor) Transcription factor 6 like 2 Transcription factor 6-like 2 transcription factor 6-like 3 Transcription factor A, mitochondrial Transcription factor A, mitochondrial Transcription factor A, mitochondrial precursor |
Images
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Fig1:
Western blot analysis of mtTFA on different lysates with Rabbit anti-mtTFA antibody (ET1705-64) at 1/1,000 dilution. Lane 1: K-562 cell lysate (20 µg/Lane) Lane 2: HL-60 cell lysate (20 µg/Lane) Lane 3: HeLa cell lysate (20 µg/Lane) Lane 4: HepG2 cell lysate (20 µg/Lane) Lane 5: MCF7 cell lysate (20 µg/Lane) Lane 6: Human brain tissue lysate (40 µg/Lane) Predicted band size: 29 kDa Observed band size: 24 kDa Exposure time: 1 minute; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-64) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling mtTFA with Rabbit anti-mtTFA antibody (ET1705-64) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-64, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig3:
Immunofluorescence analysis of paraffin-embedded human colon tissue labeling mtTFA with Rabbit anti-mtTFA antibody (ET1705-64) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-64, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig4:
Immunocytochemistry analysis of HeLa cells labeling mtTFA with Rabbit anti-mtTFA antibody (ET1705-64) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-mtTFA antibody (ET1705-64) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-mtTFA antibody (ET1705-64) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-64) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig6:
mtTFA was immunoprecipitated from 0.2 mg K-562 cell lysate with ET1705-64 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1705-64 at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/100,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: ET1705-64 IP in K-562 cell lysate Lane 3: Rabbit K-562 instead of ET1705-64 in 293T cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute; ECL: K1801 |
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