ET1703-74

TDP43 Recombinant Rabbit Monoclonal Antibody [JM51-10]

cat.: ET1703-74

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IP, IF-Cell, IF-Tissue, IHC-P, FC, IHC-Fr
Clonality:	Monoclonal
Clone number:	JM51-10

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 45 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human TDP43 aa 354-390 / 414; (2)aa 253-290 / 414.
Positive control:	HeLa cell lysate, K-562 cell lysate, Neuro-2a cell lysate, mouse spleen tissue lysate, mouse brain tissue lysate, rat spleen tissue lysate, rat brain tissue lysate, HeLa, Neuro-2a, human spleen tissue, mouse placenta tissue, human pancreas tissue, mouse hippocampus tissue, mouse cerebral cortex tissue, mouse cerebrum tissue.
Subcellular location:	Cytoplasm, Mitochondrion, Nucleus.
Recommended Dilutions: WB IHC-Fr IF-Cell IF-Tissue IHC-P FC IP	1:500-1:5,000 1:200-1:500 1:250-1:500 1:100 1:500 1:50-1:100 Use at an assay dependent concentration.
Uniprot #:	SwissProt: Q13148 Human \| Q921F2 Mouse Entrez Gene: 298648 Rat \| 325052 Zebrafish
Alternative names:	ALS10 OTTHUMP00000002171 OTTHUMP00000002172 OTTHUMP00000002173 TADBP_HUMAN TAR DNA binding protein 43 TAR DNA binding protein TAR DNA-binding protein 43 TARDBP TDP 43 TDP-43 TDP43

Images

Fig1: Western blot analysis of TDP43 on different lysates with Rabbit anti-TDP43 antibody (ET1703-74) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: Mouse spleen tissue lysate
Lane 5: Mouse brain tissue lysate
Lane 6: Rat spleen tissue lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 20 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-74) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunofluorescence analysis of frozen mouse cerebrum tissue with Rabbit anti-TDP43 antibody (ET1703-74) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1703-74, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

	Fig3: Immunocytochemistry analysis of HeLa cells labeling TDP43 with Rabbit anti-TDP43 antibody (ET1703-74) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TDP43 antibody (ET1703-74) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunocytochemistry analysis of Neuro-2a cells labeling TDP43 with Rabbit anti-TDP43 antibody (ET1703-74) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TDP43 antibody (ET1703-74) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig5: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-TDP43 antibody (ET1703-74) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-74) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-TDP43 antibody (ET1703-74) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-74) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling TDP43 with Rabbit anti-TDP43 antibody (ET1703-74). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1703-74, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
	Fig8: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling TDP43 with Rabbit anti-TDP43 antibody (ET1703-74). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1703-74, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
	Fig9: Flow cytometric analysis of TDP43 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-74, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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