PA5-144005

antibody from Invitrogen Antibodies

Targeting: YTHDC2 DKFZp564A186, FLJ10053, FLJ2194

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-144005

Provider

Invitrogen Antibodies

Product name

YTHDC2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Adding 0.2 mL of distilled water will yield a concentration of 500 µg/mL. Positive Control - WB: human Daudi whole cell, human Hela whole cell, human 293T whole cell, rat C6 whole cell, mouse NIH/3T3 whole cell. ICC/IF: SiHa cell. Flow: MCF-7 cell, SiHa cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of YTHDC2 using YTHDC2 antibody (Product # PA5-144005). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. Lane 1: human Daudi whole cell lysates; Lane 2: human Hela whole cell lysates; Lane 3: human 293T whole cell lysates; Lane 4: rat C6 whole cell lysates; Lane 5: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-YTHDC2 antigen affinity purified polyclonal antibody (Product # PA5-144005) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for YTHDC2 at approximately 175 kDa. The expected band size for YTHDC2 is at 160 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of YTHDC2 in SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. Samples were then incubated in YTHDC2 Polyclonal antibody (Product # PA5-144005) using a dilution of 5 μg/mL. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of YTHDC2 in SiHa cells (Blue line). The cells were blocked with 10% normal goat serum, and then incubated with YTHDC2 polyclonal antibody (Product # PA5-144005) (1 µg/1x10^6 cells) for 30 min at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6 cells) used under the same conditions. Unlabeled sample (Red line) was also used as a control. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C.