Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-TDP43 antibody [EPR5810] (ab109535)

Knockout Tested Rabbit recombinant monoclonal TDP43 antibody [EPR5810]. Validated in WB, IP, IHC, ICC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 3 publication(s).

Overview

  • Product name

    Anti-TDP43 antibody [EPR5810]
    See all TDP43 primary antibodies
  • Description

    Rabbit monoclonal [EPR5810] to TDP43
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human TDP43 (N terminal). The exact sequence is proprietary.
    Database link: Q13148

  • Positive control

    • WB: HAP1, HeLa, Jurkat, 293T, K562, and A431 cell lysates, Mouse and Rat brain lysates; IHC-Fr: Mouse cerebrum tissue; IHC-P: Human papillary carcinoma and glioma tissue, Mouse and Rat cerebrum tissues; ICC/IF: HAP1-TARDBP and HeLa cells; FC: K562 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109535 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/50.
Flow Cyt Use at an assay dependent concentration.
IHC-Fr 1/50.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

WB 1/1000 - 1/10000. Predicted molecular weight: 45 kDa.
IHC-P 1/100 - 1/250. antigen retrieval is recommended

Target

  • Function

    DNA and RNA-binding protein which regulates transcription and splicing. Involved in the regulation of CFTR splicing. It promotes CFTR exon 9 skipping by binding to the UG repeated motifs in the polymorphic region near the 3'-splice site of this exon. The resulting aberrant splicing is associated with pathological features typical of cystic fibrosis. May also be involved in microRNA biogenesis, apoptosis and cell division. Can repress HIV-1 transcription by binding to the HIV-1 long terminal repeat. Stabilizes the low molecular weight neurofilament (NFL) mRNA through a direct interaction with the 3' UTR.
  • Tissue specificity

    Ubiquitously expressed. In particular, expression is high in pancreas, placenta, lung, genital tract and spleen.
  • Involvement in disease

    Defects in TARDBP are the cause of amyotrophic lateral sclerosis type 10 (ALS10) [MIM:612069]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology of ALS is likely to be multifactorial, involving both genetic and environmental factors. The disease is inherited in 5-10% of the cases.
  • Sequence similarities

    Contains 2 RRM (RNA recognition motif) domains.
  • Domain

    The RRM domains can bind to both DNA and RNA.
  • Post-translational
    modifications

    Hyperphosphorylated in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU.
    Ubiquitinated in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU.
    Cleaved to generate C-terminal fragments in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU.
  • Cellular localization

    Nucleus. In patients with frontotemporal lobar degeneration and amyotrophic lateral sclerosis, it is absent from the nucleus of affected neurons but it is the primary component of cytoplasmic ubiquitin-positive inclusion bodies.
  • Information by UniProt
  • Database links

  • Alternative names

    • ALS10 antibody
    • OTTHUMP00000002171 antibody
    • OTTHUMP00000002172 antibody
    • OTTHUMP00000002173 antibody
    • TADBP_HUMAN antibody
    • TAR DNA binding protein 43 antibody
    • TAR DNA binding protein antibody
    • TAR DNA-binding protein 43 antibody
    • TARDBP antibody
    • TDP 43 antibody
    • TDP-43 antibody
    • TDP43 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: TDP43 knockout HAP1 cell lysate (40 µg)
    Lane 3: HeLa cell lysate (40 µg)
    Lane 4: Jurkat cell lysate (40 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab109535 observed at 48 kDa. Red - loading control, ab8245, observed at 37 kDa.

    Unpurified ab109535 was shown to specifically react with TDP43  when TDP43  knockout samples were used. Wild-type and TDP43  knockout samples were subjected to SDS-PAGE. Ab109535 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling TDP43 with purified ab109535 at 1/100 dilution (0.3 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TDP43 with purified ab109535 at 1/50 dilution (6.2 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • All lanes : Anti-TDP43 antibody [EPR5810] (ab109535) at 1/5000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa

  • Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling TDP43 with purified ab109535 at 1/20 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling TDP43 with Purified unpurified ab109535 at 1/50 (0.5 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling TDP43 with purified ab109535 at 1/100 dilution (0.3 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Unpurified ab109535 staining TDP43 in wild-type HAP1 cells (top panel) and TDP43 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109535 at 1μg/ml concentration and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling TDP43 with purified ab109535 at 1/100 dilution (0.3 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling TDP43 with unpurified ab109535, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human glioma.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • All lanes : Anti-TDP43 antibody [EPR5810] (ab109535) at 1/5000 dilution ((unpurified))

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TDP43 with unpurified ab109535, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse cerebrum.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • All lanes : Anti-TDP43 antibody [EPR5810] (ab109535) at 1/1000 dilution ((unpurified))

    Lane 1 : HeLa cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : A431 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 45 kDa

  •  Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling TDP43 with unpurified ab109535, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebrum.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2)

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Unpurified ab109535 at 1/100 dilution staining TARDBP in paraffin-embedded Human papillary carcinoma tissue by Immunohistochemistry.

References

This product has been referenced in:

  • Zhao L  et al. TDP-43 is Required for Mammary Gland Repopulation and Proliferation of Mammary Epithelial Cells. Stem Cells Dev N/A:N/A (2019). Read more (PubMed: 31062657) »
  • Seminary ER  et al. Modeling Protein Aggregation and the Heat Shock Response in ALS iPSC-Derived Motor Neurons. Front Neurosci 12:86 (2018). Read more (PubMed: 29515358) »
See all 4 Publications for this product

Customer reviews and Q&As

Filter by Application

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Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Dorsal Root Ganglion)
Specification
Dorsal Root Ganglion

Steven West

Verified customer

Submitted Oct 27 2017

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