Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Other assay [1]
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Validation data
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- Product number
- PA5-17232 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SIRT1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody. This antibody is not cross-reactive with other sirtuin proteins.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 21 µg/mL
- Storage
- -20°C
Submitted references Niacin Inhibits Apoptosis and Rescues Premature Ovarian Failure.
Wang S, Sun M, Yu L, Wang Y, Yao Y, Wang D
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2018;50(6):2060-2070
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2018;50(6):2060-2070
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SirT1 was performed by loading 20 µg of THP-1 whole cell lysates per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% milk in TBST for 1 hr at room temperature. The membrane was probed with a SirT1 polyclonal antibody (Product # PA5-17232) at a dilution of 1:500 overnight at °°C, washed in TBST, and probed with a HRP-conjugated goat anti-rabbit IgG at a dilution of 1:40,000 for 1 hr at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SirT1 was performed by loading 20 µL of HeLa or COS whole cell lysates per well onto a SDS-PAGE gel. Proteins were transferred to a membrane and blocked with 5% non-fat dry milk for 1 hour at room temperature. The membrane was probed with a SirT1 polyclonal antibody (Product # PA5-17232) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with a peroxidase-conjugated anti-rabbit IgG secondary antibody for 1 hour at room temperature. Detection was performed using a chemiluminescent substrate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SIRT1 Polyclonal Antibody (Product # PA5-17232) and a 110 kDa band corresponding to SIRT1 was observed in all cell lysates. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), HEK-293 (Lane 3) and NTERA-2 cl.D1 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SIRT1 was performed using 70% confluent log phase HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SIRT1 Polyclonal Antibody (Product # PA5-17232) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of SirT1 (green) HEK293T cells. Cells fixed in 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with a SirT1 polyclonal antibody (Product # PA5-17232) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of SirT1 was performed on HEK293T cells. Antigen-antibody complexes were formed by incubating 500 µg of HEK293T whole cell lysate (in 300 µL volume with 5 µL of a SirT1 polyclonal antibody (Product # PA5-17232) or a rabbit IgG (negative control) overnight at 4°C. The immune complexes were captured on 30 µL of protein G sepharose beads, washed extensively, and eluted with 6X Laemmli buffer. Samples were resolved on an 8% SDS-PAGE gel, transferred to a PVDF membrane, and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with a SirT1 polyclonal antibody (Product # PA5-17232) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with a peroxidase-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.