Anti-MDH4 | Malate dehydrogenase 4 (cytoplasmic)

(Cat#: AS15 3065)
AS15_3065
AS15_3065_1
Description
  • Immunogen: Recombinant MDH4 of Zea mays, UniProt: Q08062
  • Host: Rabbit
  • Clonality: Polyclonal
  • Purity: Serum
  • Format: Lyophilized
  • Quantity: 50 µl
  • Reconstitution: For reconstitution add 50 µl of sterile water
  • Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
  • Tested applications: Western blot (WB)
  • Recommended dilutions: 1 : 1000 (WB)
  • Expected | apparent MW: 33.5 kDa
  • Confirmed reactivity: Zea mays
  • Malate dehydrogenase 4 (cytoplasmic), (EC:1.1.1.37) display catalytic activity in conversion of (S)-malate + NAD+ = oxaloacetate + NADH.
  • Lim et al (2022). Arabidopsis guard cell chloroplasts import cytosolic ATP for starch turnover and stomatal opening. Nat Commun. 2022 Feb 3;13(1):652. doi: 10.1038/s41467-022-28263-2. PMID: 35115512; PMCID: PMC8814037.
  • Immunoprecipitation was performed by using Dynabeads ® Protein A: briefly 100 µl suspension was washed with 200 µl TTBS (Tris Buffered saline, 50 mM Tris-HCl pH 7.6 and 165 mM NaCl with 0.1% Tween 80) using the magnetic stands for concentrating the magnetic beads. After wash the beads were preincubated with 20 µl primary antibodies in 180 µl TTBS at room temperature for 30 minutes (minimum 15 minues). A first wash was followed afterwards with 200 µl TTBS and hence a real incubation with 200 µl plant extract (supernatant 20,000 x g for 3 min.), 200µl of TTBS and further 50 µl YeastBuster reagent (Novagen) containing a mixture of detergents to break and solubilize the mitochondria membrane. This incubation at room temperature was allowed to be under mild shaking to allow the beads to be in suspension. Hence supernatant was aspirated away by the use of the magnetic stand and two further washesing steps with 200 µl TTBS were performed prior mixing with 100 µl SDS-Sample buffer.Antibody is also binding to recombinant MHD4 (see image below).

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