PA5-29949

antibody from Invitrogen Antibodies

Targeting: TARDBP ALS10, TDP-43

Provider product page for PA5-29949

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Chromatin Immunoprecipitation

Other assay

Antibody data

Antibody Data
Antigen structure
References [3]
Comments [0]
Validations
Western blot [10]
Immunocytochemistry [4]
Other assay [3]

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Product number: PA5-29949 - Provider product page
Provider: Invitrogen Antibodies
Product name: TDP-43 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: 293T, A431, HeLa, HepG2, Neuro2A, GL261, NIH-3T3, BCL-1, Raw264.7, PC-12, Rat-2, Rat brain. Predicted reactivity: Mouse (95%), Rat (95%), Zebrafish (83%), Xenopus laevis (91%), Chicken (95%), Bovine (96%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.63 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

TARDBP protein structure - PA5-29949 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TARDBP using 50 µg of rat brain lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a TARDBP polyclonal antibody (Product # PA5-29949) at a dilution of 1:3000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TARDBP using 30 µg of A) A431 (B) H1299 and C) HeLa lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a TARDBP polyclonal antibody (Product # PA5-29949) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDP-43 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDP-43 Polyclonal Antibody (Product # PA5-29949) at a dilution of 1:2000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDP-43 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDP-43 Polyclonal Antibody (Product # PA5-29949) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using TDP-43 Polyclonal Antibody (Product # PA5-29949). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with TDP-43 Polyclonal Antibody (Product # PA5-29949) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TARDBP antibody detects TARDBP protein by western blot analysis. A. 30 µg Neuro2A whole cell lysate/extract. B. 30 µg GL261 whole cell lysate/extract. C. 30 µg NIH-3T3 whole cell lysate/extract. D. 30 µg BCL-1 whole cell lysate/extract. E. 30 µg Raw264.7 whole cell lysate/extract.10% SDS-PAGE. TARDBP antibody TDP-43 Polyclonal Antibody (Product # PA5-29949) dilution: 1:3,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDP-43 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDP-43 Polyclonal Antibody (Product # PA5-29949) at a dilution of 1:2000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: CRISPR-Cas9 mediated genome editing of TARDBP was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042) (Assay ID CRISPR1087594_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of TARDBP was performed by loading 30 µg of HeLa CAS9 (Lane 1) and HeLa CAS9 cells transduced with TARDBP Lentiviral sgRNA (Lane 2) whole cell extracts. The blot was probed with Anti-TDP-43 Polyclonal Antibody (Product # PA5-29949) using 1:3000 dilution and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific to TARDBP (Fig (b)). An uncharacterized band was observed at ~32 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of TDP-43 was achieved by transfecting HeLa with TDP-43 specific siRNAs (Silencer® select Product # s23830, s23829). Western blot analysis (Fig. a) was performed using Whole Cell Extract-WCL from the TDP-43 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TDP-43 Polyclonal Antibody (Product # PA5-29949, 1:3000 dilution ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to TDP-43.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-TDP-43 Polyclonal Antibody(Product # PA5-29949) and a 44kDa band corresponding to TDP-43 was observed across cell lines tested. Whole Cell Extract-WCL (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), Caco-2 (Lane 3) and K-562 (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of TARDBP in paraformaldehyde-fixed HeLa cells using a TARDBP polyclonal antibody (Product # PA5-29949) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TDP-43 Polyclonal Antibody detects TDP43 protein by immunofluorescent analysis. Sample: DIV10 rat E18 primary cortical neuron cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: TDP43 stained by TDP-43 Polyclonal Antibody (Product # PA5-29949) diluted at 1:500. Red: Tau, stained by Phospho-Tau (Ser262) Polyclonal Antibody [GT287]diluted at 1:500. Blue: Fluoroshield with DAPI .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TDP-43 Polyclonal Antibody detects TDP43 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: TDP43 stained by TDP-43 Polyclonal Antibody (Product # PA5-29949) diluted at 1:2,000. Red: phalloidin, a cytoskeleton marker, diluted at 1:200.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TDP-43 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TDP-43 Polyclonal Antibody (Product # PA5-29949) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TARDBP antibody immunoprecipitates TARDBP protein in IP experiments. IP Sample: HeLa whole cell lysate/extract A. 40 µg HeLa whole cell lysate/extract B. Control with 2 µg of preimmune rabbit IgG C. Immunoprecipitation of TARDBP protein by 2 µg of TARDBP antibody (Product # PA5-29949) 12% SDS-PAGE The immunoprecipitated TARDBP protein was detected by TARDBP antibody (Product # PA5-29949) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Neuropathologic Changes in More Advanced Cases of Chronic Traumatic Encephalopathy Widespread cortical tau deposits in superficial cortical layers and involving both gyri and sulci in a continuous pattern. Some of the superficial cortical pathology can be focal and involve the crown of the gyri with widespread white matter punctate tau deposits in the (B and D) white matter and (C) subventricular areas. Other concomitant pathologies are alpha-synuclein deposits in the shape of Lewy bodies and neurites as depicted in the (E) cingulate cortex and (F) TAR DNA-binding protein 43 deposits as shown in the dentate gyrus. Scale bar in the top left corner represents 0.1 cm in panels A and B and 0.01 cm in panels C-F.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure S5. Transcriptome-wide RNA-seq, m 6 A-seq, and RIP-seq assays in murine NK cells. (A) Volcano plots of differentially expressed genes from RNA-seq. (B) GSEA showing enrichment of E2F targets, G2/M checkpoint, and mitotic spindle hallmark gene sets in Ythdf2 DeltaNK NK cells compared with Ythdf2 WT NK cells. (C) The proportion of m 6 A peak distribution in NK cells from Ythdf2 WT and Ythdf2 DeltaNK mice. (D) GO analysis of transcripts with m 6 A peaks. (E) Density distribution of the YTHDF2-binding sites across mRNA transcriptome from RIP-seq data. (F) The proportion of YTHDF2 binding site distribution from RIP-seq data. (G) Top 10 GO clusters from GO analysis of YTHDF2 target genes from RIP-seq data. (H) Immunoblotting showing the protein levels of MDM2, TDP-43, and YTHDF2 in IL-2-expanded NK cells enriched from the spleen of Ythdf2 WT and Ythdf2 DeltaNK mice. (I and J) qPCR and immunoblotting showing the expression of Mdm2 (I) and Tardbp (J) in NK cells transfected with gene-specific or control siRNA. (K and L) IL-2-expanded NK cells were transfected with Mdm2 -specific siRNA cells under the stimulation of IL-15. 3 d later, cell proliferation and apoptosis were analyzed by Ki67 staining (K) and annexin V staining (L), respectively ( n = 3 per group). Data are shown as mean +- SD and were analyzed by unpaired two-tailed t test (I and J) or one-way ANOVA with Sidak post-test (K and L). Data are representative of at least two independent experiments. ***, P < 0.001. ES,