- MA5-15677 - Invitrogen Antibodies SIRT1 antibody

Submitted references Oxidative Stress-Induced Sirtuin1 Downregulation Correlates to HIF-1α, GLUT-1, and VEGF-A Upregulation in Th1 Autoimmune Hashimoto's Thyroiditis.

Hepp M, Werion A, De Greef A, de Ville de Goyet C, de Bournonville M, Behets C, Lengelé B, Daumerie C, Mourad M, Ludgate M, Many MC, Joris V, Craps J
International journal of molecular sciences 2021 Apr 7;22(8)

Desoxyrhapontigenin attenuates neuronal apoptosis in an isoflurane-induced neuronal injury model by modulating the TLR-4/cyclin B1/Sirt-1 pathway.

Liang F, Fu X, Li Y, Han F
AMB Express 2020 Sep 30;10(1):175

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SIRT1 was achieved by transfecting MDA-MB-231 cells with SIRT1 specific siRNAs (Silencer® select Product # s223591, s223592). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the SIRT1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with SIRT1 Monoclonal Antibody (Product # MA5-15677, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SIRT1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of MCF7 (Lane1), Jurkat (Lane 2), HeLa (Lane 3), A549 (Lane 4), NTERA-2 (Lane 5), PANC-1 (Lane 6) and HEK-293 (Lane 7). The blot was probed with SIRT1 Monoclonal Antibody (1F3) (Product # MA5-15677, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). A 110kDa band corresponding to SIRT1 was observed across the panel.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SIRT1 using SIRT1 monoclonal antibody (Product # MA5-15677) in MCF-7 (1), Jurkat (2), HeLa (3), HEK293 (4) and A549 (5) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SIRT1 was performed by loading 100 ng Cal-51 cell lysate in RIPA Lysis and Extraction buffer (Product # 89900) and Page Ruler Plus Protein Ladder (Product # 26619) onto a 30% Acrylamide/Bis gel. Proteins were transferred and membrane was blocked in 5% milk for one hour at room temperature. SIRT1 was detected at approximately 120 kDa using a SIRT1 Monoclonal Antibody (Product # MA5-15677) at a dilution of 1:1000 in 5% milk overnight at at 4°C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236). Data courtesy of Dr Wei Xu's lab at University of Wisconsin Madison.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SIRT1 was performed using 70% confluent log phase NTERA-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SIRT1 Monoclonal Antibody (1F3) (Product # MA5-15677) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization of SIRT1. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of NTERA-2 cells using SIRT1 monoclonal antibody (Product # MA5-15677) (Green). Red: actin filaments have been labeled with phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of K562 cells using SIRT1 monoclonal antibody (Product # MA5-15677) (green) and negative control (purple).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 8 Desoxyrhapontigenin ameliorates the protein expression of Sirt-1, according to immunohistochemical analysis, in the brain tissues of rats with anaesthesia-induced neuronal injury

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 In Th1-HT context, SIRT1 was downregulated while HIF-alpha was upregulated. Addition of Th1 cytokines to primary cultures of human thyrocytes induced a significant reduction of SIRT1 ( A ) and an increase of HIF-1alpha protein ( B ). Densitometric values were normalized against the beta-actin level. Results are expressed as means +- SEM from seven (SIRT1) and four (HIF-1alpha) experiments ( n = 4-7) at least in duplicate. SIRT1 protein expression ( C ) was significantly decreased in HT patients while HIF-1alpha ( D ) was increased. Densitometric values were normalized against the GAPDH level. Results are expressed as means +- SEM from five or six individual samples ( n = 5-6). * p < 0.05, *** p < 0.005 compared to controls. Representative blots are shown. Paranodular tissue from multinodular goiter patients designated as controls ( E , H ) and HT thyroid samples ( F , G , I , J ) were used to perform IHC with SIRT1 and HIF-1alpha specific antibodies. In controls ( E ), SIRT1 was detected in thyrocytes of hypofunctional follicles (+) and more intensively in active follicles (*). In HT ( F , G ), staining was less intense in altered active-like follicles ($). In control thyroids ( H ), HIF-1alpha staining was observed in the cytoplasm of the thyrocytes (+ and *). In HT thyroids ( I , J ), non-inflammatory and inflammatory zones displayed hypofunctional (+) and altered active-like ($) follicles with strong staining of HIF-1alpha in thyrocyte cytoplasm. Illustrations sho

MA5-15677

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