Anti-mtTFA TFAM Antibody Picoband™ (monoclonal, 4D9)

Product Name

Anti-mtTFA TFAM Antibody Picoband™ (monoclonal, 4D9)

View all mtTFA Antibodies

SKU/Catalog Number

M01119-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-mtTFA TFAM Antibody Picoband™ (monoclonal, 4D9) catalog # M01119-1. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-mtTFA TFAM Antibody Picoband™ (monoclonal, 4D9) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01119-1)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Monoclonal

Clone Number

4D9

Isotype

Mouse IgG2b

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human mtTFA, different from the related mouse and rat sequences by five amino acids.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M01119-1 is reactive to TFAM in Human

Applications

M01119-1 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

40 kDa

Calculated molecular weight

29.097kDa

Background of mtTFA

TFAM (Transcription factor A, mitochondrial), also known as TCF6 or TCF6L2, is a 162-amino acid protein that activates transcription of each mitochondrial DNA (mtDNA) strand by binding to an element of approximately 30 nucleotides present in both the light-strand and the heavy-strand promoters. By Southern blot analysis of restriction enzyme digests of human/Chinese hamster somatic cell hybrid lines, Milatovich et al. (1992) mapped TFAM sequences, which they called MTTF1, to 3 different chromosomes: chromosomes 10, 7p, and 11q. By PCR-based screening of a somatic cell hybrid panel and by fluorescence in situ hybridization, Scott (2007) stated that the sequences mapped to chromosomes 7p (TCF6L1) and 11q (MTTF1, or TCF6L3) are pseudogenes. Larsson et al. (1997) mapped the mouse mitochondrial transcription factor A gene (Tfam) to the central part of mouse chromosome 10. This region exhibits syntenic homology with human 10q21. Mitochondrial transcription factor A is a key activator of mitochondrial transcription in mammals. It also has a role in mitochondrial DNA replication, since transcription generates an RNA primer necessary for initiation of mtDNA replication.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 1-2μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of TFAM using anti ZO-1 antibody (M01119-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEK293 tissue lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human A549 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-TFAM antigen affinity purified polyclonal antibody (Catalog # M01119-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for TFAM at approximately 40KD. The expected band size for TFAM is at 40KD.

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Figure 2. IHC analysis of TFAM using anti-TFAM antibody (M01119-1).
TFAM was detected in paraffin-embedded section of human rectum cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-TFAM Antibody (M01119-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 3. IHC analysis of TFAM using anti-TFAM antibody (M01119-1).
TFAM was detected in paraffin-embedded section of human Breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-TFAM Antibody (M01119-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of CACO-2 cells using anti-FAM antibody (M01119-1).
Overlay histogram showing CACO-2 cells stained with M01119-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-FAM Antibody (M01119-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For TFAM (Source: Uniprot.org, NCBI)

Alternative Names

mitochondrial transcription factor A; MtTF1; mtTFA; TCF-6; TCF6L1; TCF6L2Mitochondrial transcription factor 1; TCF6TCF6L3; Transcription factor 6; Transcription factor 6-like 2 (mitochondrial transcription factor); Transcription factor 6-like 2; transcription factor A, mitochondrial TFAM MTDPS15, MTTF1, MTTFA, TCF6, TCF6L1, TCF6L2, TCF6L3 transcription factor A, mitochondrial transcription factor A, mitochondrial|mitochondrial transcription factor 1|mitochondrial transcription factor A|transcription factor 6|transcription factor 6-like 1|transcription factor 6-like 2 (mitochondrial transcription factor)|transcription factor 6-like 3

For more info on TFAM, check out the TFAM Infographic

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In this infographic, you will see the following information for TFAM: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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