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[KD Validated] NNMT Rabbit mAb (A25391)

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

Western blot analysis of lysates from wild type (WT) and NNMT knockdown (KD) HeLa cells using [KD Validated] NNMT Rabbit mAb (A25391) at 1:3000 dilution.
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: 30s.

ABclonal:Immunofluorescence - [KD Validated] NNMT Rabbit mAb (A25391)

Confocal imaging of A549 cells using [KD Validated] NNMT Rabbit mAb (A25391, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - [KD Validated] NNMT Rabbit mAb (A25391)

Immunoprecipitation of NNMT in 200 µg extracts from A549 cells using 0.5 µg [KD Validated] NNMT Rabbit mAb (A25391). Western blot analysis was performed using [KD Validated] NNMT Rabbit mAb (A25391) at 1:3000 dilution.

Overview

Product name	[KD Validated] NNMT Rabbit mAb
Catalog No.	A25391
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC65463

Background

N-methylation is one method by which drug and other xenobiotic compounds are metabolized by the liver. This gene encodes the protein responsible for this enzymatic activity which uses S-adenosyl methionine as the methyl donor.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-264 of human NNMT (NP_006160.1).
Sequence	MESGFTSKDTYLSHFNPRDYLEKYYKFGSRHSAESQILKHLLKNLFKIFCLDGVKGDLLIDIGSGPTIYQLLSACESFKEIVVTDYSDQNLQELEKWLKKEPEAFDWSPVVTYVCDLEGNRVKGPEKEEKLRQAVKQVLKCDVTQSQPLGAVPLPPADCVLSTLCLDAACPDLPTYCRALRNLGSLLKPGGFLVIMDALKSSYYMIGEQKFSSLPLGREAVEAAVKEAGYTIEWFEVISQSYSSTMANNEGLFSLVARKLSRPL
Gene ID
Swiss Prot
Synonyms	Nicotinamide N-methyltransferase
Calculated MW	30kDa
Observed MW	30kDa

Applications

Reactivity	Human
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 1:3000 - 1:12000 IF/ICC 1:200 - 1:800 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells 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 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunofluorescence Immunoprecipitation
Positive samples	HeLa
Cellular location	Cytoplasm.