3mC Recombinant Rabbit Monoclonal Antibody [PSH03-22]
cat.: HA721958
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Species independent
Applications: ELISA, IF-Cell, Dot Blot
Clonality: Monoclonal
Clone number: PSH03-22
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: 3-mc-OVA
Positive control: U-2 OS cells treated with 4mM Methyl Methanesulfonate for 1 hour.
Recommended Dilutions:
  IF-Cell
  Dot Blot

1:100
1:2,000
Images
HA721958_1.jpg Fig1: Immunocytochemistry analysis of U-2 OS cells treated with 4mM Methyl Methanesulfonate for 1 hour labeling 3mC with Rabbit anti-3mC antibody (HA721958) at 1/100 dilution.

Cells were fixed in 70% ethyl alcohol for 5 minutes at room temperature, then subjected to acid hydrolysis using 2M HCl in TBST for 30 minutes at room temperature. Permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-3mC antibody (HA721958) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
HA721958_2.jpg Fig2: Dot blot analysis of 3mC on different proteins with Rabbit anti-3mC antibody (HA721958) at 1/2,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.

Lane 1: Uridine-BSA (negative)
Lane 2: Cytidine-BSA (negative)
Lane 3: 5-Methylcytosine-BSA (negative)
Lane 4: 3-Methylcytosine-BSA (positive)
Lane 5: 5-Hydroxymethylcytidine-BSA (negative)

Proteins loading: 50ng, 25ng, 12.5ng, 6.25ng;

Blocking and dilution buffer: 5% NFDM/TBST;

Exposure time: 1 minute.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.