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Affinity Purified Rabbit Polyclonal Antibody (Pab)

AP18410c staining ALKBH5 in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (1/100) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Overlay histogram showing U-2 OS cells stained with AP18410c(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP18410c, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(1583138) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
All lanes : Anti-ALKBH5 Antibody (Center) at 1:500 dilution Lane 1: Hela whole cell lysate Lane 2: MCF-7 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 44 kDa Blocking/Dilution buffer: 5% NFDM/TBST.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, FC, E
Primary Accession	Q6P6C2
Other Accession	D3ZKD3, Q3TSG4, E1BH29, NP_060228.3
Reactivity	Human, Rat, Mouse
Predicted	Mouse, Rat, Bovine, Canine, Rabbit, Chicken
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	44256 Da
Antigen Region	302-330 aa

Additional Information
Gene ID	54890
Other Names	RNA demethylase ALKBH5, 11411-, Alkylated DNA repair protein alkB homolog 5, Alpha-ketoglutarate-dependent dioxygenase alkB homolog 5, ALKBH5, ABH5, OFOXD1
Target/Specificity	This ALKBH5 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 302-330 amino acids from the Central region of human ALKBH5.
Dilution	WB~~1:500 IHC-P~~1:100 FC~~1:25 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	ALKBH5 Antibody (Center) is for research use only and not for use in diagnostic or therapeutic procedures.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	ALKBH5 {ECO:0000303\|PubMed:23177736, ECO:0000312\|HGNC:HGNC:25996}
Function	Dioxygenase that specifically demethylates N(6)- methyladenosine (m6A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes (PubMed:23177736, PubMed:24489119, PubMed:24616105, PubMed:24778178, PubMed:34048572, PubMed:36944332, PubMed:37257451, PubMed:37369679, PubMed:40793791). Demethylates RNA by oxidative demethylation, which requires molecular oxygen, alpha-ketoglutarate and iron (PubMed:21264265, PubMed:23177736, PubMed:24489119, PubMed:24616105, PubMed:24778178). Demethylation of m6A mRNA affects mRNA processing, translation and export (PubMed:23177736, PubMed:34048572, PubMed:36944332, PubMed:37257451). Can also demethylate N(6)-methyladenosine in single-stranded DNA (in vitro) (PubMed:24616105). Required for the late meiotic and haploid phases of spermatogenesis by mediating m6A demethylation in spermatocytes and round spermatids: m6A demethylation of target transcripts is required for correct splicing and the production of longer 3'-UTR mRNAs in male germ cells (By similarity). Involved in paraspeckle assembly, a nuclear membraneless organelle, by undergoing liquid-liquid phase separation (PubMed:37369679, PubMed:37474102). Paraspeckle assembly is coupled with m6A demethylation of RNAs, such as NEAT1 non-coding RNA (PubMed:37474102). Also acts as a negative regulator of T-cell development: inhibits gamma-delta T-cell proliferation via demethylation of JAG1 and NOTCH2 transcripts (By similarity). Inhibits regulatory T-cell (Treg) recruitment by mediating demethylation and destabilization of CCL28 mRNAs (By similarity).
Cellular Location	Nucleus speckle. Nucleus, nucleoplasm. Note=Promotes formation and localizes to paraspeckles, a nuclear membraneless organelle
Tissue Location	Widely expressed, with highest expression in lung, followed by testis, pancreas, spleen and ovary