MAb to Glyceraldehyde-3-PDH Antibody (1mg)

Catalog#: OAMA01120

• Size: 1mg

• Gene Symbol: GAPDH
• Alias Symbols: Monoclonal Antibody to Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH)
• Full Name: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH)
• Protein Accession#: NP_002037.2
• Swissprot Id: P04406
• Host: Mouse. Hybridization of Sp2/0 myeloma cells with spleen cells from BALB/c mice. --- Source: Ascites
• Specificity: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH)
  Cross-reacts with GAPDH from Human, bovine, porcine, goat, rabbit and rat.
• Application: ELISA, ICC, IA, WB
• Clone: B903M
• Product Format: Purified, Liquid
• Purity: >90% pure (SDS-PAGE). Protein A chromatography
• Isotype: IgG1
  Buffer: PBS, pH 7.4
  
  Preservative: 0.1% Sodium azide
  
  Important Note: Centrifuge before opening to ensure complete recovery of vial contents.
  
  Warnings: This product contains sodium azide, which has been classified as Xn (Harmful), in European Directive 67/548/EEC in the concentration range of 0.1 - 1.0 %. When disposing of this reagent through lead or copper plumbing, flush with copious volumes
• Reconstitution and Storage: Store at 2 to 8C.
• Application Data: Immunogen: Human or rabbit GAPDH
  
  Affinity Constant: Not determined

Product Protocols: Immunoblotting Protocol with MAb to Glyceraldehyde-3-PDH Antibody (1mg) Antibody (Catalog Number: OAMA01120)

Product Protocols:
Immunoblotting Protocol with Gene Name: MAb to Glyceraldehyde-3-PDH Antibody (1mg) Antibody (Catalog Number: OAMA01120)

Catalog Number: OAMA01120

Reference:
1: Wils H, Kleinberger G, Janssens J, Pereson S, Joris G, Cuijt I, Smits V, Ceuterick-de Groote C, Van Broeckhoven C, Kumar-Singh S.TDP-43 transgenic mice develop spastic paralysis and neuronal inclusions characteristic of ALS and frontotemporal lobar degeneration.Proc Natl Acad Sci U S A.2010 Feb 23;107(8):3858-63.Epub 2010 Feb 3.PubMed PMID: 20133711; PubMed Central PMCID: PMC2840518.

Product Name: MAb to Glyceraldehyde-3-PDH Antibody (1mg)

MAb to Glyceraldehyde-3-PDH Antibody (1mg)

Species: Human

Sample description: Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively.Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis.Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC By similarity.Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.

Experiment Name: Immunoblotting

Experiment Background:
Frozen tissue was weighed and homogenized in buffer containing 10 mM Hepes, 10 mM NaCl, 1 mM KH2PO4, 5 mM NaHCO3, 5 mM EDTA, 1 mM CaCl2, 0.5 mM MgCl2 (10× vol/wt).
2.After 10 min on ice, 2.5 M sucrose (0.5× vol/wt) was added.
3.Next, tissue was homogenized and centrifuged at 6,300 × g for 10 min.
4.The supernatant was collected as cytoplasmic fraction.
5.The pellet was washed three times in TSE buffer (10 mM Tris, 300 mM sucrose, 1 mM EDTA, 0.1% Nonidet P-40, 10× vol/wt), homogenized, and centrifuged at 4,000 × g for 5 min.
6.Finally the pellet was resuspended in RIPA with 2% SDS (5× vol/wt) as the nuclear fraction.
7.Also, proteins were sequentially extracted with buffers of increasing detergent strength (hypotonic, Triton X-100, myelin floatation, sarkosyl, and urea buffers).
8.All buffers were substituted with protease inhibitors (Complete Mixture; Roche) and phosphatase inhibitors (PhosphoStop; Roche) before use.
9.The following antibodies were used: rabbit anti-TDP-43 (ProteinTech Group), human specific mouse anti-TDP-43 (against residues 205-222; Abnova), rabbit anti-C-terminal TDP-43 (C-t TDPFig 43), rabbit anti-TDP-43 phosphorylation sites 409/410 (TDP-43 p409/410), rabbit anti-cleaved caspase-3, and rabbit anti-cleaved PARP (Cell Signaling Technology).
10.Detection was performed using HRP-conjugated secondary antibodies and ECL Plus Chemiluminescent Detection System (Amersham Biosciences).
1 Data were normalized to beta -actin (Sigma), GAPDH (Meridian Life Science), or Lamin A/C (N-18; Santa Cruz Biotechnology).
Experiment step:
Data were normalized to beta -actin (Sigma), GAPDH (Meridian Life Science), or Lamin A/C (N-18; Santa Cruz Biotechnology).