Anti-YTHDF2 antibody [EPR20318] (ab220163)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20318] to YTHDF2
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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Product name
Anti-YTHDF2 antibody [EPR20318]
See all YTHDF2 primary antibodies -
Description
Rabbit monoclonal [EPR20318] to YTHDF2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IP HumanWB MouseRatHumanRecombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type mESC, HEK-293T, HT-1080 treated with 10µM MG-132 for 4 hours, K562, HEK-293 and HeLa whole cell lysate. Human brain, mouse and rat testis tissue lysate; GST-tagged human YTHDF2 recombinant protein. IP: HepG2 cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20318 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution
Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
Lane 2 : YTHDF2 knockout mESC whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 62 kDa
Exposure time: 48 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
The wild-type and YTHDF2 knockout cell lysates were kindly provided by an anonymous collaborator.
ab220163 was shown to specifically react with YTHDF2 in wild-type mESC cells as signal was lost in YTHDF2 knockout cells. Wild-type and YTHDF2 knockout samples were subjected to SDS-PAGE. ab220163 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
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ab220163 Immunoprecipitating YTHDF2 in human HepG2 whole cell lysate. 10 µg of cell lysate was incubated with primary antibody (1/30). For western blotting ab220163 (1/1000) was used to confirm successful immunoprecipitation. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at (1/5000). For lane 3 Rabbit monoclonal IgG (ab172730) was used instead of ab220163.
In panel A, the lysate was stored at -80oC prior to the IP and incubation with the primary antibody was carried out overnight. In panel B, lysate was freshly made and used in the IP test immediately to minimize protein degradation. The incubation time was also shortened from overnight to 2h.
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All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/5000 dilution
Lane 1 : Untreated HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg
Lane 2 : HT-1080 treated with 10µM MG-132 (ab141003) for 4 hours, whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 62 kDa
Exposure time: 103 secondsBlocking/diluting buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution
Lane 1 : Human brain tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : Rat testis tissue lysate
Lane 4 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 5 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 2-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 62 kDaBlocking/diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane 1: 3 minutes; Lane 2: 92 seconds; Lane 3: 37 seconds; Lane 4: 8 seconds; Lane 5: 15 seconds.
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All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution
Lane 1 : GST-tagged human YTHDF1 recombinant protein, 20ng
Lane 2 : GST-tagged human YTHDF2 recombinant protein, 20ng
Lane 3 : GST-tagged human YTHDF3 recombinant protein, 20ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 62 kDa
Exposure time: 3 secondsBlocking/diluting buffer and concentration: 5% NFDM/TBST.
The YTHDF recombinant proteins were kindly provided by an anonymous collaborator.
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