Imprint® Monoclonal Anti-5-hydroxymethylcytosine antibody produced in mouse

Name	Imprint® Monoclonal Anti-5-hydroxymethylcytosine antibody produced in mouse
Supplier	Sigma-Aldrich
Catalog	SAB4800049
Prices	$525.00
Sizes	50 µg
Host	Mouse
Applications	MeDIP ELISA
Species Reactivities	Human, Mouse
Description	Mouse
Conjugate	unconjugated
Supplier Page	Shop

Product images

<B>Immunoblotting</B><BR/>Anti-5-hydroxymethylcytosine : <B>Cat. No. SAB4800049</B>: Dot blot analysis of the 5-hydroxymethylcytosine mouse monoclonal antibody with cytosine (C), methylcytosine (mC) and hydroxymethylcytosine (hmC) PCR controls. 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-base) of hmC (1), mC (2) and C (3) PCR controls were spotted on a membrane. The membrane was incubated with 2 μg/mL (1:500 dilution) of the antibody. The membrane was exposed for 30 seconds.

Immunoblotting
Anti-5-hydroxymethylcytosine : Cat. No. SAB4800049: Dot blot analysis of the 5-hydroxymethylcytosine mouse monoclonal antibody with cytosine (C), methylcytosine (mC) and hydroxymethylcytosine (hmC) PCR controls. 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-base) of hmC (1), mC (2) and C (3) PCR controls were spotted on a membrane. The membrane was incubated with 2 μg/mL (1:500 dilution) of the antibody. The membrane was exposed for 30 seconds.

<B>ChIP Results Chart</B><BR/>Anti-5-hydroxymethylcytosine: <B>Cat. No. SAB4800049</B>: Hydroxymethylated DNA IP (hMeDIP) was performed using the mouse monoclonal antibody directed against 5-hydroxymethylcytosine. The IgG isotype antibodies from mouse was used as negative control. The DNA was sonicated with Bioruptor® to have DNA fragments of 300-500 bp. 1 μg of human HeLa cells DNA were spiked with non-methylated, methylated,and hydroxymethylated PCR fragments. The IP minuted material has been analysed by qPCR using the primer pair specific for the 3 different control sequences. The obtained results show that the mouse monoclonal for 5-hmC is highly specific for this base modification, no IP with non-methylated or methylated C bases containing fragments.

ChIP Results Chart
Anti-5-hydroxymethylcytosine: Cat. No. SAB4800049: Hydroxymethylated DNA IP (hMeDIP) was performed using the mouse monoclonal antibody directed against 5-hydroxymethylcytosine. The IgG isotype antibodies from mouse was used as negative control. The DNA was sonicated with Bioruptor® to have DNA fragments of 300-500 bp. 1 μg of human HeLa cells DNA were spiked with non-methylated, methylated,and hydroxymethylated PCR fragments. The IP minuted material has been analysed by qPCR using the primer pair specific for the 3 different control sequences. The obtained results show that the mouse monoclonal for 5-hmC is highly specific for this base modification, no IP with non-methylated or methylated C bases containing fragments.

<B>ELISA</B><BR/>Anti-5-hydroxymethylcytosine : <B>Cat. No. SAB4800049</B>: Determination of the 5-hydroxymethylcytosine (5-hmC) mouse monoclonal antibody titer. An ELISA was performed using a serial dilution of the antibody in antigen-coated wells. The antigen used was KHL coupled to 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.

ELISA
Anti-5-hydroxymethylcytosine : Cat. No. SAB4800049: Determination of the 5-hydroxymethylcytosine (5-hmC) mouse monoclonal antibody titer. An ELISA was performed using a serial dilution of the antibody in antigen-coated wells. The antigen used was KHL coupled to 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.