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Catalog Number | orb654390 |
---|---|
Category | Antibodies |
Description | TIF1 gamma/TRIM33 Antibody (monoclonal, 8I8) |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 8I8 |
Tested applications | FC, ICC, IF, IHC, WB |
Reactivity | Human |
Isotype | Mouse IgG2b |
Immunogen | E.coli-derived human TIF1 gamma recombinant protein (Position: M1001-K1127). Human TIF1 gamma shares 96.1% amino acid (aa) sequence identity with mouse TIF1 gamma. |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 150 kDa |
UniProt ID | Q9UPN9 |
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 TIF1 gamma/TRIM33 using anti-TIF1 gamma/TRIM33 antibody (orb654390). 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 MCF-7 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human K562 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-TIF1 gamma/TRIM33 antigen affinity purified monoclonal antibody (Catalog # orb654390) 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 # orb90502) with Tanon 5200 system. A specific band was detected for TIF1 gamma/TRIM33 at approximately 150KD. The expected band size for TIF1 gamma/TRIM33 is at 150KD.
IHC analysis of TIF1 gamma/TRIM33 using anti TIF1 gamma/TRIM33 antibody (orb654390). TIF1 gamma/TRIM33 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 mouse anti-TIF1 gamma/TRIM33 Antibody (orb654390) 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 # orb90443) with DAB as the chromogen.
IHC analysis of TIF1 gamma/TRIM33 using anti TIF1 gamma/TRIM33 antibody (orb654390). TIF1 gamma/TRIM33 was detected in paraffin-embedded section of human rectal 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-TIF1 gamma/TRIM33 Antibody (orb654390) 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 # orb90443) with DAB as the chromogen.
IF analysis of TIF1 gamma/TRIM33 using anti-TIF1 gamma/TRIM33 antibody (orb654390). TIF1 gamma/TRIM33 was detected in immunocytochemical section of HeLa 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 mouse anti-TIF1 gamma/TRIM33 Antibody (orb654390) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse 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 HL-60 cells using anti-TIF1 gamma/TRIM33 antibody (orb654390). Overlay histogram showing HL-60 cells stained with orb654390 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-TIF1 gamma/TRIM33 Antibody (orb654390, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 | |
Monoclonal | |
Unconjugated |
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