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 with MB-071 (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.

Immunocytochemistry/Immunofluorescence: Lactate Dehydrogenase Antibody [NB600-861] - Tissue: HeLa cells. Fixation: fixed for 5 min in 1:1 MeTOH:Acetone, blocked with incubated with blocking buffer for 15 min. Antigen retrieval: not required. Primary antibody: Lactate Dehydrogenase antibody at 1:200 for 1 h at RT. Secondary antibody: DyLight 488 conjugated Streptavidin antibody at 1:10,000 for 30 min at RT. Staining: Lactate Dehydrogenase as green fluorescent signal.

Immunocytochemistry/Immunofluorescence: Lactate Dehydrogenase Antibody [NB600-861] - Analysis of Biotin conjugate of NB600-861. Tissue: HeLa cells. Fixation: fixed for 5 min in 1:1 MeTOH:Acetone, blocked with incubated with blocking buffer for 15 min. Antigen retrieval: not required. Primary antibody: Lactate Dehydrogenase antibody at 1:2

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

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: Lactate Dehydrogenase [Rabbit Muscle].
• 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: Ion exchange chromatography

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000 - 1:5000
  • ELISA 1:2000 - 1:10000
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:2000
  • Immunoprecipitation 1:100
• Application Notes: This antibody has been tested for use in ELISA and by western blot. Expect a band approximately 36 kDa in size corresponding - LDH western blotting in the appropriate cell lysate or extract.

Reactivity Notes

Cross-reacts with Rabbit. Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Ion exchange chromatography

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

• 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]
• 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).
• 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.

Bioinformatics

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