Lactate Dehydrogenase Antibody

Western Blot: Lactate Dehydrogenase Antibody [NB600-861] - Analysis using HRP conjugate of NB600-861. Lane 1-4: HeLa cell extracts cytoplasmic fraction (CF). Lane 5-8: HeLa cell extracts nuclear fraction (NF). Load: 30 ug per lane. Primary antibody: LDH antibody at 1:400 for overnight at 4C. Secondary antibody: IRDye800 secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO/TBST overnight at 4C. Predicted/Observed size: 36.6 kDa, 36 kDa for LDH. Other band(s): None.

Immunocytochemistry/Immunofluorescence: Lactate Dehydrogenase Antibody [NB600-861] - Analysis of HRP conjugate of NB600-861. Tissue: HeLa cells. Fixation: fixed for 5 min in 1:1 MeTOH:Acetone, blocked (preservative free) for 15 min. Antigen retrieval: not required. Primary antibody: Lactate Dehydrogenase antibody at 1:200 for

Western Blot: Lactate Dehydrogenase Antibody [NB600-861] - Analysis using Biotin conjugate of NB600-861. Lane 1: Lactate Dehydrogenase (Rabbit Muscle). Load: 50 ng per lane. Primary antibody: Goat anti-Lactate Dehydrogenase Antibody Biotin Conjugated at 1:1,000 overnight at 4C. Secondary antibody: HRP streptavidin secondary antibody at 1:40,000 for 30 min at RT. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 36.5 kDa, 36.5 kDa for Lactate Dehydrogenase.

Western Blot: Lactate Dehydrogenase Antibody [NB600-861] - Analysis using Biotin conjugate of NB600-861. Lane 1-4: HeLa cell extracts cytoplasmic fraction (CF). Lane 5-8: HeLa cell extracts nuclear fraction (NF). Load: 30 ug per lane. Primary antibody: LDH antibody at 1:400 for overnight at 4C. Secondary antibody: IRDye800 secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO/TBST overnight at 4C. Predicted/Observed size: 36.6 kDa, 36 kDa for LDH. Other band(s): None.

Western Blot: Lactate Dehydrogenase Antibody [NB600-861] - Analysis using HRP conjugate of NB600-861. Lane 1: Lactate Dehydrogenase (Rabbit Muscle). Load: 50 ng per lane. Primary antibody: Goat anti-Lactate Dehydrogenase Antibody Peroxidase Conjugated at 1:1,000 overnight at 4C. Secondary antibody: n/a. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 36.5 kDa, 36.5 kDa for Lactate Dehydrogenase.

• Reactivity: Hu, Rb
• Applications: WB, ELISA, ICC/IF, IP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Lactate Dehydrogenase Antibody Summary

• Description: This antibody is purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above.
• Immunogen: This Lactate Dehydrogenase Antibody was prepared from whole goat serum produced by repeated immunizations with a full length Lactate Dehydrogenase protein isolated from rabbit muscle. (Uniprot: P13491)
• Localization: Cytoplasmic
• Specificity: Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum as well as purified and partially purified Lactate Dehydrogenase [Rabbit Muscle].
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: LDHB
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:2000-1:10000
  • Immunocytochemistry/Immunofluorescence 1:200
  • Immunoprecipitation 1:100
  • Western Blot 1:1000-1:5000
• Application Notes: Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 36 kDa in size corresponding to LDH western blotting in the appropriate cell lysate or extract.
• Theoretical MW: 36.56 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

BLAST analysis was used to determine that cross reactivity is suggested for both muscle and heart isoforms (LDH-A and LDH-B) from most mammalian species

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Multi-step
• Reconstitution Instructions: Reconstitute with 100 ul deionized water (or equivalent)

Notes

This antibody is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum as well as purified and partially purified Lactate Dehydrogenase [Rabbit Muscle]

Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Alternate Names for Lactate Dehydrogenase Antibody

• EC 1.1.1
• EC 1.1.1.27
• lactate dehydrogenase B
• LDH heart subunit
• LDH-B
• LDH-H
• L-lactate dehydrogenase B chain
• Renal carcinoma antigen NY-REN-46
• TRG-5

Background

Lactate dehydrogenase is also known as L-lactate dehydrogenase A chain, LDHA, LDH muscle subunit and LDH-M. Two isozymes of LDH occur in mammals, LDH-M and LDH-H which come together to form a homotetramer of 36 kDa subunits. Every LDH molecule consists of four subunits, where each subunit is either H or M (based on their electrophoretic properties.) There are, therefore, five LDH isotypes: LDH-1 (4H) -in the heart, LDH-2 (3H1M) -in the reticuloendothelial system, LDH-3 (2H2M) -in the lungs, LDH-4 (1H3M) -in the kidneys and LDH-5 (4M) -in the liver and striated muscle. Usually LDH-2 is the predominant form in the serum. An LDH-1 level higher than the LDH-2 level (a "flipped pattern") suggests myocardial infarction (damage to heart tissues releases heart LDH, which is rich in LDH-1, into the bloodstream). In general, LDH is often used as a marker of tissue breakdown. LDH shows a cytoplasmic localization.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Lactate Dehydrogenase Antibody (NB600-861)(3)

Publications using NB600-861

• Koukourakis MI et al. Lactate dehydrogenase-5 (LDH-5) overexpression in non-small-cell lung cancer tissues is linked to tumour hypoxia, angiogenic factor production poor prognosis Br J Cancer 2003 [PMID: 12942121]
  
  Details:
  This citation used the HRP version of this antibody.
• Koukourakis MI et al. Lactate dehydrogenase-5 (LDH-5) overexpression in non-small-cell lung cancer tissues is linked to tumour hypoxia, angiogenic factor production poor prognosis Br J Cancer Jun 25 1905
  
  Details:
  This publication used the HRP conjugated form of this antibody (Cat# NB600-863).
• Crawford RM et al. M-LDH serves as a sarcolemmal K(ATP) channel subunit essential for cell protection against ischemia. EMBO J 21:3936-48. 2002 [PMID: 12145195]

Bioinformatics

• Gene Symbol: LDHB
• Entrez:
  • 3945(Human)
• Uniprot:
  • P07195()