- Gene Symbol:
- NCBI Gene Id:
- Official Gene Full Name:
- NADH dehydrogenase (ubiquinone) Fe-S protein 1, 75kDa (NADH-coenzyme Q reductase)
- Protein Name:
- NADH-ubiquinone oxidoreductase 75 kDa subunit, mitochondrial
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Alias Symbols:
- CI-75Kd, MGC26839, PRO1304, CI-75k
- Replacement Item:
- This antibody may replace item sc-271387 from Santa Cruz Biotechnology.
- Description of Target:
- The protein encoded by this gene belongs to the complex I 75 kDa subunit family. Mammalian complex I is composed of 45 different subunits. It locates at the mitochondrial inner membrane. This protein has NADH dehydrogenase activity and oxidoreductase activity. It transfers electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone. This protein is the largest subunit of complex I and it is a component of the iron-sulfur (IP) fragment of the enzyme. It may form part of the active site crevice where NADH is oxidized. Mutations in this gene are associated with complex I deficiency.
- Protein Size (# AA):
- Molecular Weight:
- Affinity Purified
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express NDUFS1.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express NDUFS1.
- The immunogen is a synthetic peptide directed towards the middle region of human NDUFS1
- Species Reactivity:
- Cow, Dog, Goat, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Zebrafish
- Predicted Homology Based on Immunogen Sequence:
- Cow: 100%; Dog: 93%; Goat: 100%; Guinea Pig: 93%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Zebrafish: 79%
- Complete computational species homology data:
- Anti-NDUFS1 (ARP56609_P050)
- Peptide Sequence:
- Synthetic peptide located within the following region: TPPGLAREDWKIIRALSEIAGMTLPYDTLDQVRNRLEEVSPNLVRYDDIE
- Product Format:
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
- Reconstitution and Storage:
- For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
- Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
- Protein Interactions:
- SUMO2; HSFY1; UBC; MDM2; RNF2; KDM1A; FBXO6; VCAM1; BTK; Htt; NDUFS7; RHOT2; NDUFA12; TMEM126B; MRPL13; STX12; SEC24C; SDHB; RPL27; RAB5B; OPA1; NDUFV3; NDUFV2; NDUFS8; NDUFV1; NDUFS2; NDUFB10; NDUFB9; NDUFB5; NDUFA9; CPT1A; ATP5C1; ATP5B; CUL3; SIRT7; MY
- Blocking Peptide:
- For anti-NDUFS1 (ARP56609_P050) antibody is Catalog # AAP56609 (Previous Catalog # AAPP39316)
- Printable datasheet for anti-NDUFS1 (ARP56609_P050) antibody
Researcher: Dr. Bruno Guigas, PhD, Dept. of Molecular Cell Biology, Leiden University Medical Center
Application: Western blotting
Species + Tissue/Cell type: Human, mouse, rat liver and muscle
How many ug's of tissue/cell lysate run on the gel:
1: 30 ug Human liver
2: 30 ug Rat liver
3: 30 ug Mouse (wild-type) liver
4: 30 ug Mouse [AMPK alpha 1/2(-/-)] liver
5: 30 ug Human muscle
6: 30 ug Rat muscle
7: 30 ug Mouse muscle
Primary antibody dilution: 1:1000
Secondary antibody dilution: 1:10000
1. How do Aviva's reagents play a role in your experimental goals?
Reliable antibodies are crucial for us.
2. How many different experimental trials were conducted using the antibody sample?
1 set of WB experiment was conducted.
3. What type of experimental sample are you using and how did you prepare it?
With have used 6 or 7 different samples (30 ug per lane):
1. Human liver
2. Rat liver
3. Wild-type mouse liver
4. AMPKa1+2-/- mouse liver
5. Human muscle
6. Rat muscle
7. Mouse muscle
A small piece (~30mg) of every organ was homogenized by Ultra-Turax (22.000 rpm; 2x5 sec) in a 6:1 (v/w) ratio of ice-cold buffer containing: 50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.6), 50 mM sodium fluoride, 50 mM potassium chloride, 5 mMsodium pyrophosphate, 1 mMethylenediaminetetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 5 mM β-glycerophosphate, 1 mM sodium vanadate, 1 mMdithiothreitol, 1% nonylphenoxypolyethoxylethanol (Tergitol-type NP40) and protease inhibitors cocktail (Complete, Roche, Mijdrecht, The Netherlands). Homogenates were centrifuged (13,200 rpm; 15 min, 4C) and the protein content of the supernatant was determined using a bicinchoninic acid (BCA) protein assay kit (BCA Protein Assay Kit, Thermo Scientific Pierce Protein Research Products, Rockford, US).
4. What applications did you test the antibody in? Please include dilutions of the primary and secondary reagents.
The antibodies were tested for Western Blot application (see above)
Dilution primary antibodies: 1/1000 for all
Dilution secondary antibodies: 1/10000 for all
5. What controls were used in your experiment? Please include your positive control.
No specific control samples but the proteins of interest are either expressed in all or in some of the samples used.
6. For IHC, what antigen retrieval method did you use?
7. How did you store the antibody after re-suspension?
All the antibodies were stored at -20C.
8. Please provide the protocol for your application procedure. Please be as detailed as possible.
Proteins (30 ug) were separated by either 12.5 or 10% SDS-PAGE followed by transfer to a polyvinylidene fluoride transfer membrane. Membranes were blocked for 1 h at room temperature in tris-buffered saline tween-20 buffer with 5% non-fat dry milk followed by an overnight incubation with antibodies. Blots were then incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature. Bands were visualized by enhanced chemiluminescence.
9. Please provide an image of your results. How does it compare to your previous images?
See the attachment image.
10. How would you rate this antibody on a scale from 1-5? Why?
5 very good (only one major specific band)
11. Would you use this antibody in future experiments?
The NDUFS1 is quite specific and would certainly be interesting for forthcoming experiments.
12. Do you believe the information about the reagent on Aviva's website is correct?
15. If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?