TDP-43 (N-terminal) Recombinant antibody

• Featured Product
• KD/KO Validated

TDP-43 (N-terminal) Uni-rAb^TM Recombinant Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human, mouse, rat and More (1)

Applications

WB, IHC, IF-P, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

11N20

Reviews (1)

Cat no : 80001-1-RR

Synonyms

ALS10, TAR DNA binding protein, TAR DNA binding protein 43, TARDBP, TDP 43, TDP43, tdp-43

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Validation Data Gallery

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  WB analysis using 80001-1-RR

  Various cell lysates were subjected to SDS PAGE followed by western blot with 80001-1-RR (TDP43 antibody) at dilution of 1:35000 incubated at room temperature for 1.5 hours.

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  WB analysis of HAP1 using 80001-1-RR

  HAP1 (WT and TARDBP KO) lysates prepared with NP-40 buffer, 50 μg protein loaded. 80001-1-RR incubated at 1:1000 at 4°C overnight in 5% milk in TBST. Ponceau stained transfers shown on right. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  WB analysis of HeLa using 80001-1-RR

  WB result of TDP-43 antibody (80001-1-RR; 1:30000; incubated at room temperature for 1.5 hours) with sh-Control and sh-TDP-43 transfected HeLa cells.

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  IHC staining of rat brain using 80001-1-RR

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 80001-1-RR (TDP-43 antibody) at dilution of 1:2000 (under 4x lens).

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  IHC staining of spinal cord slides from ALS patients using 80001-1-RR

  IHC results of TDP43 rabbit recombinant antibody (80001-1-RR, 1:20000) with the spinal cord slides from ALS patients. IHC experiment was done with Ventana automatic staining system and Optiview DAB detection kit with heat-induced epitope retrieval (boiling for 32 min in Tris-EDTA based solution CC1 buffer, Ventana). Fig from the lab of Dr. Neumann.

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  IHC staining of frontal cortex from FTLD-TDP type B using 80001-1-RR

  IHC results of TDP43 rabbit recombinant antibody (80001-1-RR, 1:20000) with the frontal cortex from FTLD-TDP type B patients. IHC experiment was done with Ventana automatic staining system and Optiview DAB detection kit with heat-induced epitope retrieval (boiling for 32 min in Tris-EDTA based solution CC1 buffer, Ventana). Fig from the lab of Dr. Neumann.

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  IHC staining of human gliomas using 80001-1-RR

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 80001-1-RR (TDP-43 antibody) at dilution of 1:1500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human gliomas using 80001-1-RR

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 80001-1-RR (TDP-43 antibody) at dilution of 1:1500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 80001-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 80001-1-RR (TDP-43 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of rat brain using 80001-1-RR

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using 80001-1-RR (TDP-43 antibody) at dilution of 1:150 and CoraLite488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HAP1 using 80001-1-RR

  HAP1 WT cells (yellow outline) and TARDBP KO cells (red outline) labelled with a green or a far red fluorescence dye, respectively. Cells fixed with 4% PFA and stained with 80001-1-RR at 1:400 plus DAPI. Bars = 10 μm. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  FC experiment of HeLa using 80001-1-RR

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human TDP-43 (80001-1-RR, Clone:11N20) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HeLa cells, HAP1 cells, K-562 cells, Neuro-2a cells, C6 cells
• Positive IHC detected in: human gliomas tissue, spinal cord slides from ALS patients, frontal cortex from FTLD-TDP type B, mouse brain tissue, rat brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: rat brain tissue, HAP1 cells
• Positive FC (Intra) detected in: HeLa cells
• Positive FC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:4000
• Immunofluorescence (IF)-P: IF-P : 1:300-1:1200
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below
• IF: See 1 publications below
• IP: See 1 publications below

Product Information

80001-1-RR targets TDP-43 (N-terminal) in WB, IHC, IF-P, FC (Intra), IP, ELISA applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: fusion protein
• Full Name: TAR DNA binding protein
• Calculated Molecular Weight: 43 kDa
• Observed Molecular Weight: 43 kDa
• GenBank Accession Number: BC001487
• Gene Symbol: TDP-43
• Gene ID (NCBI): 23435
• RRID: AB_2882933
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The TARDBP gene encodes the TDP-43 protein, initially found to repress HIV-1 transcription by binding TAR DNA. TDP-43 has since been shown to bind RNA as well as DNA, and have multiple functions in transcriptional repression, translational regulation and pre-mRNA splicing. For instance, it is reported to regulate alternate splicing of the CTFR gene. In 2006 Neumann et al. found that hyperphosphorylated, ubiquitinated and/or cleaved forms of TDP-43, collectively known as pathological TDP-43, play a major role in the disease mechanisms of ubiquitin-positive, tau- and alpha-synuclein-negative frontotemporal dementia (FTLD-U) and in amyotrophic lateral sclerosis (ALS). Proteintech's 80001-1-RR is a rabbit recombinant TDP-43 antibody recognizing N-terminal TDP-43. It recognizes the intact 43 kDa protein as well as all posttranslationally modified and truncated forms in multiple applications. Various forms of TDP-43 exist, including 18-35 kDa of cleaved C-terminal fragments, 45-50 kDa phospho-protein, 55 kDa glycosylated form, 75 kDa hyperphosphorylated form, and 90-300 kDa cross-linked form. (PMID: 17023659, 19823856, 21666678, 22193176) Recently TDP-43 has been reported to be overexpressed in triple negative breast cancer (TNBC) and it may be a potential target for TNBC diagnosis and drug design. (PMID: 29581274).

80001-1-RR can be used in WB, IHC, IHF. For the ICC (IF with cells) experiment, the results will be better with ethanol fixed cell and dilution 1:50 or 1:100. Another antibody with CatNo. 80002-1-RR is recommended for IF or ICC experiment.

Publications

Species	Application	Title
human	WB,IP,IF	F1000Res; The identification of high-performing antibodies for TDP-43 for use in Western Blot, immunoprecipitation and immunofluorescence; Authors - Donovan Worrall

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Silvia (Verified Customer) (02-20-2020)

Good antibody. I have used it to stain both Ethanol and NBF fixed human Control and FTLD-TDP brain section and spinal cord from ALS cases by IHC. I have tested either citrate buffer or FA as antigen retrieval solutions, and in both conditions nicely detected the normal nuclear TDP-43 as well as cytoplasmic inclusions specially in spinal cord form ALS cases . The dilutions that worked better showing high specificity and low background ranged from 5,000-10,000.

• Applications: Immunohistochemistry,
• Primary Antibody Dilution: 1:5,000 1: 10,000
• Cell Tissue Type: human cortex and spinal cord

TDP-43 (N-terminal) Antibody Immunohistochemistry, validation (1:5,000 1: 10,000 dilution) in human cortex and spinal cord (Cat no:80001-1-RR)