LDHA Protein, Human (His)

LDHA protein catalyzes the stereospecific interconversion of pyruvate and lactate, concomitantly exchanging NADH and NAD(+). LDHA Protein, Human (His) is the recombinant human-derived LDHA protein, expressed by E. coli , with N-6*His labeled tag.

LDHA protein catalyzes the simultaneous and stereospecific interconversion of pyruvate and lactate, accompanied by the reciprocal interconversion of NADH and NAD(+).

Measured by its ability to reduce pyruvate to lactate. The specific activity is > 70000 pmol/min/μg, as measured under the described conditions.

Materials
Assay buffer: 25 mM Tris, 100 mM NaCl, pH 7.5
LDHA Protein, Human (His) (HY-P70225)
beta-NADH (diluted to 20 mM with 0.1 M sodium borate), pH 9.0
sodium pyruvate (HY-W015913)

Procedure
1. Dilute Human LDHA to 0.4 ng/uL using assay buffer.
2. Substrate mixture: A substrate mixture containing 1.6 mM beta-NADH and 4 mM sodium pyruvate was prepared using assay buffer.
3. Experimental group: 50 μL protein + 50 μL substrate mixture.
Blank group: 50 μL assay buffer + 50 μL substrate mixture.
4. Plates were read in kinetic mode at 340 nm (absorbance) for 5 minutes.
5. Calculate specific activity:

Human

E. coli

N-6*His

P00338-1 (M1-F332)

3939  [NCBI]

Full Length of Isoform-1

MATLKDQLIYNLLKEEQTPQNKITVVGVGAVGMACAISILMKDLADELALVDVIEDKLKGEMMDLQHGSLFLRTPKIVSGKDYNVTANSKLVIITAGARQQEGESRLNLVQRNVNIFKFIIPNVVKYSPNCKLLIVSNPVDILTYVAWKISGFPKNRVIGSGCNLDSARFRYLMGERLGVHPLSCHGWVLGEHGDSSVPVWSGMNVAGVSLKTLHPDLGTDKDKEQWKEVHKQVVESAYEVIKLKGYTSWAIGLSVADLAESIMKNLRRVHPVSTMIKGLYGIKDDVFLSVPCILGQNGISDLVKVTLTSEEEARLKKSADTLWGIQKELQF

Approximately 34.0-38.0 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Solution

Supplied as a 0.22 μm filtered solution of 20 mM Tris-HCl, 500 mM NaCl, 5% Trehalose, 5% Mannitol, 0.01% Tween80, 1 mM EDTA, 50% Glycerol, pH 8.0 or 20 mM Tris-HCl, 100 mM NaCl, pH 8.0, 20% Glycerol.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.