Product Description

The affinity purified rabbit polyclonal anti-methylated lysine antibody was developed using a technique unique to ImmuneChem. The methylated lysine antibodies are affinity purified using N-methyl (epsilon amino group) lysine on agarose as the affinity matrix. The purified antibody was conjugated to horse radish peroxidase (HRP) via reductive amination. Direct labelling of primary anti-MeK will avoid the use of secondary antibodies therefore eliminating the interference of the secondary antibody-conjugates. The antibodies could be utilized for detection, quantization and isolation of proteins with MeK residues.

Western blot analysis of the methylated protein pattern in whole HeLa cells with ICP0502.

A. Signal of methylated proteins

B. Signal of the methylated proteins was competitively suppressed by 50 µg/mL of methylated BSA

C. Signal of the methylated proteins was competitively suppressed by 10 µg/mL of methylated BSA

Species

Rabbit

Formulation

500 µg/mL in HRP stabilizing buffer with 50% glycerol

Immunogen

Methylated KLH conjugates

Purification

Methyl-lysine and di-methyl lysine on agarose

Conjugation

Reductive amination; HRP/Ab molar ratio - 10:1

Specificity

Anti-MeK peroxidase conjugates recognize proteins methylated on their lysine residues (mono and di-methyllysine). They do not cross react with acetylated proteins.

Applications

ELISA; WB

Scientific Description

MeK is a conserved post-translational modification and is an important biochemical process for many protein-protein interactions. It is found in many proteins, for example calmodulin, cytochrome C, chromosomal proteins, histones and non-histones as well as neural storage body proteins. It has been suggested that methylation of lysine plays an important role in gene silencing. ^[1-3]

Storage & Stability

Product is stable for several weeks at 4°C. For extended storage, store product at -20°C. Expiration date is one year from date of shipping if stored properly.

Product Specific References

1. J. Biol. Chem. 2003. 278: 18346-18352. doi.10.1074/jbc.M300890200

2. J. Biol. Chem. 2002. 277: 34655-34657. doi:10.1074/jbc.C200433200

3. J. Biol. Chem. 2002. 277: 11621-11624. doi:10.1074/jbc.C200045200

Product Description

The affinity purified rabbit polyclonal anti-Di-methylated lysine antibodies are developed using a technique unique to ImmuneChem. The Di-methylated lysine antibodies are affinity purified using N-Di-methyl (epsilon amino group) lysine on agarose as the affinity matrix. The purified antibodies are conjugated to horse radish peroxidase (HRP). The antibody-HRP conjugates could be utilized for detection, quantization and localization of proteins with Di-methylated lysine residues.

Western blot analysis of the di-methylated proteins from swine spleen cell lysate with (B) and without (A) the presence of methylated BSA, using anti-di-methyl lysine antibody HRP conjugates (anti-MeK HRP, ICP0702)

Species

Rabbit

Formulation

250 µg/mL in Tris-phosphate buffer with 1% BSA and 50% glycerol

Immunogen

Di-methylated KLH conjugates

Purification

The antibody is  immunoaffinity purified with epsilon N-Di-methyl lysine on agarose.

Conjugation

Reductive amination with antibody/HRP molar ratio = 1:1

Specificity

The antibody recognizes proteins or peptides with di-methyl lysine residues. There is no cross reaction with acetylated proteins. The cross reactivity with mono-methyl lysine has not been tested.

Applications

ELISA, WB

Scientific Description

Protein lysine methylation (MeK) is a conserved post-translational modification and is an important biochemical process for many protein-protein interactions. It is found in many proteins, for example, calmodulin, cytochrome C, chromosomal proteins, histones, non-histones as well as neural storage body proteins. It has been suggested that di-methylation of lysine plays an important role in gene silencing.^[1]

Storage & Stability

Product is stable for several weeks at 4°C. For extended storage, store product at -20°C. Expiration date is one year from date of shipping if stored properly. Do not aliquot. Shake thoroughly before use.

Product Specific References

1.  1. Mol. BioSyst. 2013. 9: 2231-2247. doi: 10.1039/C3MB00009E

Product Description

This rabbit polyclonal antibody is isolated and affinity purified from the anti-serum against chemically methylated protein antigen. The affinity purified trimethylated lysine antibodies are affinity absorbed with N-(epsilon) mono- and di-methylated lysine. The purified antibody is conjugated to horse radish peroxidase (HRP) via reductive amination. Direct labelling of primary antibodies will avoid the use of secondary antibodies therefore eliminating the effects of the secondary antibody-conjugates. The antibodies could be utilized for detection and quantization of proteins with N-trimethylated lysine residues.

Western blot analysis of trimethylated protein from human melanoma cells (MMRU) with ICP0602

A: Competitive inhibition with N-(epsilon) Trimethyl-lysine (50µg/mL)

B: Without competitive inhibition with N-(epsilon) trimethyl lysine

Western blot analysis of trimethylated protein from the acid extract of calf spleen with ICP0602 

C: Competitive inhibition with N-(epsilon) trimethyl lysine (50µg/mL)

D: Without competitive inhibition with N-(epsilon) trimethyl lysine

Formulation

500 µg/mL in HRP stabilizing buffer with 50% glycerol

Immunogen

Methylated KLH conjugates

Purification

N-(epsilon) trimethyllysine on agarose and immunoaffinity absorbed with N-(epsilon) methylated lysine and N-(epsilon) dimethylated lysine on agarose

Conjugation

Reductive amination; HRP/Ab molar ratio - 2:1

Specificity

Recognizes proteins with trimethylation on lysine residues (N-epsilon); antibody does not cross react with acetylated proteins or mono- and dimethylated proteins.

Species Tested

Human, rat, mouse, bovine and pig

Applications

ELISA (1:5000); WB (1:1000)

Scientific Description

In the biosynthetic pathway of 1-carnitine, epsilon-N-trimethyllysine hydroxylase is the first enzyme that catalyzes the formation of beta-hydroxy-N-epsilon-trimethyllysine from epsilon-N-trimethyllysine. This reaction is dependent on alpha ketoglutarate, Fe²⁺, and oxygen.

Storage & Stability

Product is stable for several weeks at 4°C. For extended storage, and store product at -20°C. Expiration date is three years from date of shipping if stored properly.

Product Specific References

1. J. Biol. Chem. 2001. 36(276): 33512-33517.  doi:10.1074/jbc.M105929200