Imprint® Anti-5-hydroxymethylcytosine antibody produced in rabbit

Name	Imprint® Anti-5-hydroxymethylcytosine antibody produced in rabbit
Supplier	Sigma-Aldrich
Catalog	SAB4800051
Prices	$450.00
Sizes	100 µl
Host	Rabbit
Clonality	Polyclonal
Applications	MeDIP DB ELISA
Species Reactivities	Human, Mouse
Description	Rabbit Polyclonal
Conjugate	unconjugated
Supplier Page	Shop

Product images

<B>Immunoblotting</B><BR/>Anti-5-hydroxymethylcytosine: <B>Cat. No. SAB4800051</B>: Dot blot analysis was performed, to determine the antibody specificity, using C, mC and hmC containing PCR products. 100 to 4 ng, equivalent of 5 to 0.2 pmol of C-bases, were spotted on a membrane, Amersham Hybond-N+. The membrane was incubated with the Rabbit 5-hydroxymethylcytosine polyclonal antibody (dilution 1:200). The membranes were exposed for 30 seconds.

Immunoblotting
Anti-5-hydroxymethylcytosine: Cat. No. SAB4800051: Dot blot analysis was performed, to determine the antibody specificity, using C, mC and hmC containing PCR products. 100 to 4 ng, equivalent of 5 to 0.2 pmol of C-bases, were spotted on a membrane, Amersham Hybond-N+. The membrane was incubated with the Rabbit 5-hydroxymethylcytosine polyclonal antibody (dilution 1:200). The membranes were exposed for 30 seconds.

<B>ChIP Results Chart</B><BR/>Anti-5-hydroxymethylcytosine: <B>Cat. No. SAB4800051</B>: Hydroxymethylated DNA IP, hMeDIP, was performed using the Rabbit polyclonal antibody directed against 5-hydroxymethylcytosine. 1 μg of human HeLa cells DNA was spiked with non-methylated, methylated, and hydroxymethylated fragments. The IP’d material has been analyzed by qPCR using primer pairs specific for the 3 different control sequences. The obtained results show that the Rabbit polyclonal 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. SAB4800051: Hydroxymethylated DNA IP, hMeDIP, was performed using the Rabbit polyclonal antibody directed against 5-hydroxymethylcytosine. 1 μg of human HeLa cells DNA was spiked with non-methylated, methylated, and hydroxymethylated fragments. The IP’d material has been analyzed by qPCR using primer pairs specific for the 3 different control sequences. The obtained results show that the Rabbit polyclonal 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. SAB4800051</B>: Determination of the 5-hmC antibody titer. An ELISA was performed using a serial dilution of the Rabbit polyclonal antibody directed against 5-hmC in antigen coated wells. The antigen used was BSA coupled to the 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1: 3,500.

ELISA
Anti-5-hydroxymethylcytosine: Cat. No. SAB4800051: Determination of the 5-hmC antibody titer. An ELISA was performed using a serial dilution of the Rabbit polyclonal antibody directed against 5-hmC in antigen coated wells. The antigen used was BSA coupled to the 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1: 3,500.