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Catalog Number | orb654470 |
---|---|
Category | Antibodies |
Description | Transketolase/TKT Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human Transketolase/TKT recombinant protein (Position: M1-A116). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 4μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 68 kDa |
UniProt ID | P29401 |
Storage | 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. |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). 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 lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: human placenta tissue lysates, Lane 4: human Jurkat whole cell lysates, Lane 5: human HT1080 whole cell lysates, Lane 6: human HepG2 whole cell lysates, Lane 7: human SW620 whole cell lysates, Lane 8: human Raji whole cell lysates, Lane 9: rat liver tissue lysates, Lane 10: rat RH35 whole cell lysates, Lane 11: mouse HEPA1-6 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 rabbit anti-Transketolase/TKT antigen affinity purified polyclonal antibody (Catalog # orb654470) 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-rabbit 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 # orb90503) 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.
IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). Transketolase/TKT was detected in paraffin-embedded section of human mammary 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 rabbit anti-Transketolase/TKT Antibody (orb654470) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.
IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). Transketolase/TKT was detected in paraffin-embedded section of human mammary 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 rabbit anti-Transketolase/TKT Antibody (orb654470) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.
IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). Transketolase/TKT was detected in paraffin-embedded section of mouse brain 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 rabbit anti-Transketolase/TKT Antibody (orb654470) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.
IHC analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). Transketolase/TKT was detected in paraffin-embedded section of rat brain 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 rabbit anti-Transketolase/TKT Antibody (orb654470) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.
IF analysis of Transketolase/TKT using anti-Transketolase/TKT antibody (orb654470). Transketolase/TKT was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL rabbit anti-Transketolase/TKT Antibody (orb654470) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of THP-1 cells using anti-Transketolase/TKT antibody (orb654470). Overlay histogram showing THP-1 cells stained with orb654470 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Transketolase/TKT Antibody (orb654470, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of U87 cells using anti-Transketolase/TKT antibody (orb654470). Overlay histogram showing U87 cells stained with orb654470 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Transketolase/TKT Antibody (orb654470, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
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