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Product Info Summary
| SKU: | M01119-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-mtTFA TFAM Antibody Picoband® (monoclonal, 4D9)
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01119-1 is reactive to TFAM in Human
Observed Molecular Weight
40 kDa
Calculated molecular weight
29.1 kDa
Background of TFAM
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.
Application & Images
Applications
M01119-1 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee
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
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HEK293 tissue, human K562 whole cell, human Caco-2 whole cell, human Raji whole cell, human A549 whole cell,
IHC: human rectum cancer tissue, human Breast cancer tissue
FCM: CACO-2 cell
Validation Images & Assay Conditions
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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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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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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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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). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-mtTFA TFAM Antibody Picoband® (monoclonal, 4D9) (M01119-1)
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4 Customer Q&As for Anti-mtTFA TFAM Antibody Picoband® (monoclonal, 4D9)
Question
We have been able to see staining in human lymphocyte. What should we do? Is anti-mtTFA antibody (monoclonal, 4D9) supposed to stain lymphocyte positively?
Verified Customer
Verified customer
Asked: 2019-11-15
Answer
From what I have seen in literature lymphocyte does express TFAM. From what I have seen in Uniprot.org, TFAM is expressed in female gonad, hepatoma, tongue, lymphocyte, cervix carcinoma, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have TFAM expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 18669648
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Lymphocyte, Pubmed ID: 1610904
Tongue, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-11-15
Question
My team were happy with the WB result of your anti-mtTFA antibody (monoclonal, 4D9). However we have been able to see positive staining in lymphocyte mitochondrion using this antibody. Is that expected? Could you tell me where is TFAM supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-10-15
Answer
Based on literature, lymphocyte does express TFAM. Generally TFAM expresses in mitochondrion. Regarding which tissues have TFAM expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 18669648
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Lymphocyte, Pubmed ID: 1610904
Tongue, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-10-15
Question
Our lab used your anti-mtTFA antibody (monoclonal, 4D9) for Flow Cytometry on cervix carcinoma a few years ago. I am using human, and We intend to use the antibody for WB next. Our lab want to know about examining cervix carcinoma as well as female gonad in our next experiment. Could you please give me some suggestion on which antibody would work the best for WB?
Verified Customer
Verified customer
Asked: 2019-07-05
Answer
I looked at the website and datasheets of our anti-mtTFA antibody (monoclonal, 4D9) and I see that M01119-1 has been validated on human in both Flow Cytometry and WB. Thus M01119-1 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-07-05
Question
We are currently using anti-mtTFA antibody (monoclonal, 4D9) M01119-1 for human tissue, and we are satisfied with the Flow Cytometry results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on dog tissues as well?
P. Jackson
Verified customer
Asked: 2013-02-07
Answer
The anti-mtTFA antibody (monoclonal, 4D9) (M01119-1) has not been tested for cross reactivity specifically with dog tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-02-07
