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[KD Validated] DBT Rabbit mAb (A25001)

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] DBT Rabbit mAb (A25001) at 1:6000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from wild type (WT) and DBT knockdown (KD) 293F cells using [KD Validated] DBT Rabbit mAb (A25001) at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

ABclonal:Immunohistochemistry - [KD Validated] DBT Rabbit mAb (A25001)

Immunohistochemistry analysis of DBT in paraffin-embedded human kidney tissue using [KD Validated] DBT Rabbit mAb (A25001) at a dilution of 1:200 (40x lens).High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunoprecipitation - [KD Validated] DBT Rabbit mAb (A25001)

Immunoprecipitation of DBT in 300 µg extracts from Hep G2 cells using 3 µg [KD Validated] DBT Rabbit mAb (A25001). Western blot analysis was performed using [KD Validated] DBT Rabbit mAb (A25001) at 1:5000 dilution.

Overview

Product name	[KD Validated] DBT Rabbit mAb
Catalog No.	A25001
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC60670

Background

The branched-chain alpha-keto acid dehydrogenase complex (BCKD) is an inner-mitochondrial enzyme complex involved in the breakdown of the branched-chain amino acids isoleucine, leucine, and valine. The BCKD complex is thought to be composed of a core of 24 transacylase (E2) subunits, and associated decarboxylase (E1), dehydrogenase (E3), and regulatory subunits. This gene encodes the transacylase (E2) subunit. Mutations in this gene result in maple syrup urine disease, type 2. Alternatively spliced transcript variants have been described, but their biological validity has not been determined.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 62-247 of DBT (NP_001909.4).
Sequence	GQVVQFKLSDIGEGIREVTVKEWYVKEGDTVSQFDSICEVQSDKASVTITSRYDGVIKKLYYNLDDIAYVGKPLVDIETEALKDSEEDVVETPAVSHDEHTHQEIKGRKTLATPAVRRLAMENNIKLSEVVGSGKDGRILKEDILNYLEKQTGAILPPSPKVEIMPPPPKPKDMTVPILVSKPPVF
Gene ID
Swiss Prot
Synonyms	E2; E2B; BCATE2; BCKADE2; BCKAD-E2; BCKDH-E2; BCOADC-E2; [KD Validated] DBT
Calculated MW	53kDa
Observed MW	54KD/54kDa

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 1:2000 - 1:10000 IHC-P 1:50 - 1:200 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
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 Immunohistochemistry Immunoprecipitation
Positive samples	293F, Hep G2, Mouse brain
Cellular location	cytoplasm, cytosol, microtubule cytoskeleton, mitochondrial alpha-ketoglutarate dehydrogenase complex, mitochondrial matrix, mitochondrial nucleoid, mitochondrion