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Antikörper

5-Hydroxymethylcytosine (5-hmC) Recombinant Mouse monoclonal Antibody

Produktgrößen
100μg
£689,00
HA610209-100UG
Über dieses Produkt
SKU:
HA610209
Anwendung:
Western Blot, IHC-Paraffin, Dot Blot, ELISA, MeDIP
Buffer:
PBS (pH7.4).
translate.label.attr.clone:
PSH06-08
Klonalität:
Monoclonal
Konjugat:
Non-conjugated
Host:
Mouse
Immunogen:
Recombinant protein fused with 5-Hydroxymethylcytosine.
Isotyp:
IgG1
Physischer Zustand:
Liquid
Reinheit:
Protein A affinity purified.
Versandbedingungen:
Blue Ice
Lagerbedingungen:
Store at +4[o]C after thawing. Aliquot store at -20[o]C. Avoid repeated freeze / thaw cycles.
Hersteller:
Huabio
Typ:
Antibodies: Monoclonal Antibody
Western blot analysis of 5-Hydroxymethylcytosine (5-hmC) on different conjugations with Mouse anti-5-Hydroxymethylcytosine (5-hmC) antibody (HA610209) at 1/1;000 dilution. Lane 1: Uridine-BSA (negative) Lane 2: Cytidine-BSA (negative) Lane 3: 5-Methylcytosine-BSA (negative) Lane 4: 3-Methylcytosine-BSA (negative) Lane 5: 5-Hydroxymethylcytidine-BSA (positive) Lysates/proteins at 2 μg/Lane. Predicted band size: 66 kDa Observed band size: 66 kDa Exposure time: 6 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA610209) at 1/1;000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50;000 dilution was used for 1 hour at room temperature.
Competitive ELISA analysis of 5-hmc was performed by coating wells of a 96-well plate with 50 µl per well of 5-hmc-BSA diluted in carbonate/bicarbonate buffer; at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed; blocked with 1%BSA blocking buffer; and incubated with 100 µl per well of 5-hmc monoclonal antibody at concentration of 1 µg/mL with serial diluted 5-hmc and its analogs starting from a concentration of 10;000ng/ml to 0.61ng/mL for 1 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1/35;000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 8 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm. This antibody demonstrates high specificity for 5-hmc and little or no crossreactivity to it analogs.
Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-5-Hydroxymethylcytosine (5-hmC) antibody (HA610209) at 1/200 dilution. The section was denatured in 2M HCl for 15 minutes at room temperature; then neutralized in 0.1M Tris-HCl (pH 8.3) for 10 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature; washed with ddH2O and PBS; and then probed with the primary antibody (HA610209) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-5-Hydroxymethylcytosine (5-hmC) antibody (HA610209) at 1/200 dilution. The section was denatured in 2M HCl for 15 minutes at room temperature; then neutralized in 0.1M Tris-HCl (pH 8.3) for 10 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature; washed with ddH2O and PBS; and then probed with the primary antibody (HA610209) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.