Western Blot (WB)
(Figure 1. Western blot analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).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 A549 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: rat liver tissue lysatesLane 5: rat RH35 whole cell lysatesLane 6: mouse liver tissue 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- Transketolase/TKT antigen affinity purified monoclonal antibody (Catalog # MBS1754008) at 0. 25 μg/mL overnight at 4 degree 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 # MBS176445) with Tanon 5200 system. A specific band was detected for Transketolase/TKT at approximately 68KD. The expected band size for Transketolase/TKT is at 68KD. )
Mouse Transketolase/TKT Monoclonal Antibody | anti-TKT antibody
Anti-Transketolase/TKT Antibody (monoclonal, 2I3)
IHC-P: 1-2ug/ml|Human|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).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 A549 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: rat liver tissue lysatesLane 5: rat RH35 whole cell lysatesLane 6: mouse liver tissue 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- Transketolase/TKT antigen affinity purified monoclonal antibody (Catalog # MBS1754008) at 0. 25 μg/mL overnight at 4 degree 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 # MBS176445) with Tanon 5200 system. A specific band was detected for Transketolase/TKT at approximately 68KD. The expected band size for Transketolase/TKT is at 68KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).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 A549 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: rat liver tissue lysatesLane 5: rat RH35 whole cell lysatesLane 6: mouse liver tissue 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- Transketolase/TKT antigen affinity purified monoclonal antibody (Catalog # MBS1754008) at 0. 25 μg/mL overnight at 4 degree 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 # MBS176445) with Tanon 5200 system. A specific band was detected for Transketolase/TKT at approximately 68KD. The expected band size for Transketolase/TKT is at 68KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human liver 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human liver 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human lung 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human lung 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human pancreatic 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human pancreatic 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human thyroid 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in paraffin-embedded section of human thyroid 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 2μg/ml mouse anti-Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 6. IF analysis of Transketolase/TKT using anti- Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 6. IF analysis of Transketolase/TKT using anti- Transketolase/TKT antibody (MBS1754008).Transketolase/TKT was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- Transketolase/TKT Antibody (MBS1754008) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 7. Flow Cytometry analysis of THP-1 cells using anti-Transketolase/TKT antibody (MBS1754008).Overlay histogram showing THP-1 cells stained with MBS1754008 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Transketolase/TKT Antibody (MBS1754008, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
Flow Cytometry (FC/FACS)
(Figure 7. Flow Cytometry analysis of THP-1 cells using anti-Transketolase/TKT antibody (MBS1754008).Overlay histogram showing THP-1 cells stained with MBS1754008 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Transketolase/TKT Antibody (MBS1754008, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
Flow Cytometry (FC/FACS)
(Figure 8. Flow Cytometry analysis of A549 cells using anti-Transketolase/TKT antibody (MBS1754008).Overlay histogram showing A549 cells stained with MBS1754008 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Transketolase/TKT Antibody (MBS1754008, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
Flow Cytometry (FC/FACS)
(Figure 8. Flow Cytometry analysis of A549 cells using anti-Transketolase/TKT antibody (MBS1754008).Overlay histogram showing A549 cells stained with MBS1754008 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Transketolase/TKT Antibody (MBS1754008, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
2. Boyle, L., Wamelink, M. M. C., Salomons, G. S., Roos, B., Pop, A., Dauber, A., Hwa, V., Andrew, M., Douglas, J., Feingold, M., Kramer, N., Saitta, S., Retterer, K., Cho, M. T., Begtrup, A., Monaghan, K. G., Wynn, J., Chung, W. K. Mutations in TKT are the cause of a syndrome, including short stature, developmental delay, and congenital heart defects. Am. J. Hum. Genet. 98: 1235-1242, 2016.
3. Coy, J. F., Dubel, S., Kioschis, P., Thomas, K., Micklem, G., Delius, H., Poustka, A. Molecular cloning of tissue-specific transcripts of a transketolase-related gene: implications for the evolution of new vertebrate genes. Genomics 32: 309-316, 1996.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a thiamine-dependent enzyme which plays a role in the channeling of excess sugar phosphates to glycolysis in the pentose phosphate pathway. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Apr 2012]
Uniprot Description
TKT: Catalyzes the transfer of a two-carbon ketol group from a ketose donor to an aldose acceptor, via a covalent intermediate with the cofactor thiamine pyrophosphate. Homodimer. Belongs to the transketolase family.
Protein type: Transferase; EC 2.2.1.1; Carbohydrate Metabolism - pentose phosphate pathway
Chromosomal Location of Human Ortholog: 3p14.3
Cellular Component: nucleoplasm; peroxisome; cytosol; nucleus; vesicle
Molecular Function: transketolase activity; protein homodimerization activity; metal ion binding; cofactor binding
Biological Process: pentose-phosphate shunt; regulation of growth; carbohydrate metabolic process; energy reserve metabolic process; glyceraldehyde-3-phosphate biosynthetic process; pentose-phosphate shunt, non-oxidative branch; pathogenesis; xylulose biosynthetic process
Disease: Wernicke-korsakoff Syndrome
Research Articles on TKT
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Product Notes
The TKT tkt (Catalog #AAA1754008) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-Transketolase/TKT Antibody (monoclonal, 2I3) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Transketolase/TKT can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM). WB: 0.1-0.25ug/ml|Human, Mouse, Rat| IHC-P: 1-2ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the TKT tkt for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Transketolase/TKT, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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