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[KD Validated] METTL3 Rabbit mAb (A26858)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using [KD Validated] METTL3 Rabbit mAb (A26858) at 1:5000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.

Western blot analysis of lysates from Rat brain using [KD Validated] METTL3 Rabbit mAb (A26858) at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

Western blot analysis of lysates from wild type (WT) and METTL3 knockdown (KD) 293T cells using [KD Validated] METTL3 Rabbit mAb (A26858) at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 15s.

ABclonal:Immunohistochemistry - [KD Validated] METTL3 Rabbit mAb (A26858)

Immunohistochemistry analysis of paraffin-embedded Human thyroid cancer tissue using [KD Validated] METTL3 Rabbit mAb (A26858) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunoprecipitation - [KD Validated] METTL3 Rabbit mAb (A26858)

Immunoprecipitation of METTL3 from 300 µg extracts of HepG2 cells was performed using 0.5 µg of [KD Validated] METTL3 Rabbit mAb (A26858). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KD Validated] METTL3 Rabbit mAb (A26858) at a dilution of 1 : 1000.

Immunoprecipitation of METTL3 from 300 µg extracts of HepG2 cells was performed using 0.5 µg of METTL3 Rabbit mAb (A26858). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using METTL3 Rabbit mAb (A26858) at a dilution of 1:1000.

ABclonal:Chromatin Immunoprecipitation - [KD Validated] METTL3 Rabbit mAb (A26858)

Chromatin immunoprecipitation was performed with 15 μg of cross-linked chromatin from MCF7, using 3 μg of [KD Validated] METTL3 Rabbit mAb (A26858) and Rabbit IgG isotype control (AC042). The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.

Overview

Product name	[KD Validated] METTL3 Rabbit mAb
Catalog No.	A26858
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC66900

Background

This gene encodes the 70 kDa subunit of MT-A which is part of N6-adenosine-methyltransferase. This enzyme is involved in the posttranscriptional methylation of internal adenosine residues in eukaryotic mRNAs, forming N6-methyladenosine.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-156 of human METTL3 (NP_062826.2).
Sequence	MSDTWSSIQAHKKQLDSLRERLQRRRKQDSGHLDLRNPEAALSPTFRSDSPVPTAPTSGGPKPSTASAVPELATDPELEKKLLHHLSDLALTLPTDAVSICLAISTPDAPATQDGVESLLQKFAAQELIEVKRGLLQDDAHPTLVTYADHSKLSAM
Gene ID
Swiss Prot
Synonyms	M6A; IME4; Spo8; MT-A70; hMETTL3
Calculated MW	64kDa
Observed MW	75kDa/70KDa/70kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:5000 - 1:10000 IHC-P 1:200 - 1:2000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ChIP 5μg antibody for 10μg-15μg of Chromatin ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunoprecipitation
Positive samples	293T, Hep G2, Rat thymus, Mouse brain, NIH/3T3, HepG2, 293T, Rat brain
Cellular location	Nucleus speckle.