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[KD Validated] TFEB Rabbit mAb (A25981)

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 lysates from RAW 264.7 cells using [KD Validated] TFEB Rabbit mAb (A25981) 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: 45s.

Western blot analysis of lysates from wild type (WT) and TFEB knockdown (KD) 293T cells using [KD Validated] TFEB Rabbit mAb (A25981) 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: 90s.

ABclonal:Immunohistochemistry - [KD Validated] TFEB Rabbit mAb (A25981)

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue using [KD Validated] TFEB Rabbit mAb (A25981) 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.

Immunohistochemistry analysis of paraffin-embedded Human liver tissue using [KD Validated] TFEB Rabbit mAb (A25981) 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:Immunofluorescence - [KD Validated] TFEB Rabbit mAb (A25981)

Confocal imaging of A20 cells using [KD Validated] TFEB Rabbit mAb (A25981, dilution 1:300) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Chromatin Immunoprecipitation - [KD Validated] TFEB Rabbit mAb (A25981)

Chromatin immunoprecipitation was performed with 15 μg of cross-linked chromatin from Raji cells treated by Torin-1 (250nM, 5h), using 5 μg of [KD Validated] TFEB Rabbit mAb (A25981) 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] TFEB Rabbit mAb
Catalog No.	A25981
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC68094

Background

Enables DNA-binding transcription factor activity; enzyme binding activity; and transcription cis-regulatory region binding activity. Involved in several processes, including cellular response to amino acid starvation; lysosome localization; and positive regulation of autophagy. Located in cytosol; lysosomal membrane; and nucleoplasm.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 50-173 of human TFEB (NP_009093.1).
Sequence	TPAINTPVHFQSPPPVPGEVLKVQSYLENPTSYHLQQSQHQKVREYLSETYGNKFAAHISPAQGSPKPPPAASPGVRAGHVLSSSAGNSAPNSPMAMLHIGSNPERELDDVIDNIMRLDDVLGY
Gene ID
Swiss Prot
Synonyms	TCFEB; BHLHE35; ALPHATFEB
Calculated MW	44kDa/53kDa
Observed MW	65-70kDa

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 1:1000 - 1:2000 IHC-P 1:200 - 1:800 IF/ICC 1:200 - 1:400 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements. ChIP 5μg antibody for 10μg-15μg of Chromatin
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 Immunofluorescence
Positive samples	RAW 264.7, 293T
Cellular location	cytoplasm, cytosol, extracellular exosome, microtubule cytoskeleton, mitochondrial inner membrane, mitochondrial intermembrane space, mitochondrial matrix, mitochondrial nucleoid, mitochondrion, nucleus, Cytoplasm, Nucleus.