Antibody data
- Antibody Data
- Antigen structure
- References [4]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
- Other assay [4]
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- Product number
- PA5-21677 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NDUFS4 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, mouse brain, rat heart.
- Concentration
- 1 mg/mL
Submitted references Mitochondrial calcium uniporter stabilization preserves energetic homeostasis during Complex I impairment.
cAMP/PKA Signaling Modulates Mitochondrial Supercomplex Organization.
Enhancing natriuretic peptide signaling in adipose tissue, but not in muscle, protects against diet-induced obesity and insulin resistance.
Activation of mTORC1 is essential for β-adrenergic stimulation of adipose browning.
Balderas E, Eberhardt DR, Lee S, Pleinis JM, Sommakia S, Balynas AM, Yin X, Parker MC, Maguire CT, Cho S, Szulik MW, Bakhtina A, Bia RD, Friederich MW, Locke TM, Van Hove JLK, Drakos SG, Sancak Y, Tristani-Firouzi M, Franklin S, Rodan AR, Chaudhuri D
Nature communications 2022 May 19;13(1):2769
Nature communications 2022 May 19;13(1):2769
cAMP/PKA Signaling Modulates Mitochondrial Supercomplex Organization.
Signorile A, Pacelli C, Palese LL, Santeramo A, Roca E, Cocco T, De Rasmo D
International journal of molecular sciences 2022 Aug 25;23(17)
International journal of molecular sciences 2022 Aug 25;23(17)
Enhancing natriuretic peptide signaling in adipose tissue, but not in muscle, protects against diet-induced obesity and insulin resistance.
Wu W, Shi F, Liu D, Ceddia RP, Gaffin R, Wei W, Fang H, Lewandowski ED, Collins S
Science signaling 2017 Jul 25;10(489)
Science signaling 2017 Jul 25;10(489)
Activation of mTORC1 is essential for β-adrenergic stimulation of adipose browning.
Liu D, Bordicchia M, Zhang C, Fang H, Wei W, Li JL, Guilherme A, Guntur K, Czech MP, Collins S
The Journal of clinical investigation 2016 May 2;126(5):1704-16
The Journal of clinical investigation 2016 May 2;126(5):1704-16
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western Blot using NDUFS4 Polyclonal Antibody (Product # PA5-21677). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 12% SDS PAGE. NDUFS4 Polyclonal Antibody (Product # PA5-21677) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NDUFS4 Polyclonal Antibody detects NDUFS4 protein by western blot analysis. A. 50 µg Rat heart lysate/extract. 12% SDS-PAGE. NDUFS4 Polyclonal Antibody (Product # PA5-21677) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
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- Western Blot using NDUFS4 Polyclonal Antibody (Product # PA5-21677). Sample (30 µg whole cell lysate). A: 293T. 12% SDS PAGE. NDUFS4 Polyclonal Antibody (Product # PA5-21677) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of NDUFS4 was achieved by transfecting HEK-293 cells with NDUFS4 specific siRNAs (Silencer® select Product # s9396, s9395). Western blot analysis (Fig. a) was performed using membrane extracts from the NDUFS4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with NDUFS4 Polyclonal Antibody (Product # PA5-21677, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 NDUFS4. A non-specific band was also observed at ~30 kDa with the above testing conditions.
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- Western blot analysis was performed on whole extracts (30 µg lysate) of HEK 293 (Lane 1), tissue extracts of Mouse Heart (Lane 2), Rat Heart (Lane 3) and Mouse Brain (Lane 4). The blot was probed with Anti- NDUFS4 Polyclonal Antibody (Product # PA5-21677, 1:2000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 20 kDa band corresponding to NDUFS4 was detected in the cell line and tissues tested.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Immunofluorescent analysis of NDUFS4 in methanol-fixed HeLa cells using a NDUFS4 polyclonal antibody (Product # PA5-21677) at a 1:100 dilution.
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- Invitrogen Antibodies (provider)
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- Immunofluorescence analysis of methanol-fixed HeLa, using NDUFS4 antibody (Product # PA5-21677) at 1:100 dilution.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescence analysis of NDUFS4 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with NDUFS4 Rabbit Polyclonal Antibody (Product # PA5-21677) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing Mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
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- Immunohistochemical analysis of paraffin-embedded human ovarian cancer, using NDUFS4 (Product # PA5-21677) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
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- Fig. 5. Glucose uptake and expression of thermogenesis markers are increased in the BAT of HFD-fed Nprc AKO mice. ( A ) Glucose infusion rate of Nprc AKO ( n = 10) and Nprc fl/fl ( n = 8) mice during a hyperinsulinemic-euglycemic clamp experiment performed at 12 weeks of HFD feeding. ( B and C ) Rates of glucose disposal ( R d ) (B) and endogenous glucose appearance (Endo R a ) (C) in Nprc AKO ( n = 10) and Nprc fl/fl ( n = 8) mice under basal and clamp states. ( D ) Rate of glucose uptake ( R g ) in the BAT, iWAT, gWAT, gastrocnemius (GA), superficial vastus lateralis (SVL), soleus (SO), heart, and brain of Nprc AKO ( n = 10) and Nprc fl/fl ( n = 8) mice at the end of the clamp experiment. ( E ) Western blot analysis of thermogenic and mitochondrial proteins in the BAT of Nprc AKO and Nprc fl/fl mice after 12 weeks on HFD. Blots are representative of three independent experiments. COX4, cytochrome c oxidase subunit 4; CYTOC, cytochrome c; NDUFS4, NADH (reduced form of nicotinamide adenine dinucleotide) dehydrogenase ubiquinone iron-sulfur protein 4; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( F ) Immunostaining and quantification of UCP1 in the BAT of Nprc AKO ( n = 2) and Nprc fl/fl ( n = 2) mice after 12 weeks on HFD, as described in Materials and Methods. Scale bar, 200 mum. ( G ) qRT-PCR for the expression of genes coding thermogenic, mitochondrial, fatty acid oxidation (FAOx) markers, and BAT-derived adipokines in the BAT of Nprc AKO ( n = 14) and Nprc fl/fl ( n
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- Isoproterenol increased supercomplexes containing complex I, complex III and complex IV. The H9c2 cell cultures were incubated for 30 min in absence (CTRL) or in the presence of isoproterenol (ISO). Mitochondrial solubilized proteins were separated by 1D-BNE/2D-SDS-PAGE and transferred into nitrocellulose membrane, followed by immunoblotting analysis with specific antibodies. ( A ) Representative images of immunoblotting with antibody against NDUFS4. ( B ) Representative images of immunoblotting with antibodies against Core II and beta-ATP synthase. ( C ) Representative images of immunoblotting with antibody against Cox IV. ( A - C ) the histograms represent the percentage of ADU of isoproterenol-treated cells (ISO) with respect to untreated cells (CTRL) of immuno-revealed spots in free complexes (CxI, CxIII, CxIV) and in supercomplexes (SCs). The values are the means +- SD of three different experiments. (ISO vs. CTRL, *** p < 0.001, Student's t -test). For more details, see under Materials and Methods.