Anti-TDP43 antibody [2E2-D3]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TDP43 antibody [2E2-D3] (AB57105)

TARDBP antibody (ab57105) used in immunofluorescence at 10ug/ml on HeLa cells.

This image was generated using the ascites version of the product.

• Flow Cyt

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Flow Cytometry - Anti-TDP43 antibody [2E2-D3] (AB57105)

Overlay histogram showing HeLa cells stained with ab57105 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57105, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using the ascites version of the product.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TDP43 antibody [2E2-D3] (AB57105)

ab57105 staining TDP43 in human leiomyosarcoma tissue. Antibody concentration 3μg/ml.

This image was generated using the ascites version of the product.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TDP43 antibody [2E2-D3] (AB57105)

ab57105 staining TDP43 in HeLa cells. Antibody concentration 10μg/ml.

This image was generated using the ascites version of the product.

• WB

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Western blot - Anti-TDP43 antibody [2E2-D3] (AB57105)

TDP43 antibody (ab57105) at 1ug/lane + A-431 cell lysate at 25ug/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-TDP43 antibody [2E2-D3] (ab57105)

Predicted band size: 44 kDa

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• WB

Supplier Data

Western blot - Anti-TDP43 antibody [2E2-D3] (AB57105)

ab57105 staining TDP43 overexpressed in the HEK293 cell line (cotransfected with TDP43 validated Chimera siRNA) (lane 1) or non-tranfected control (lane 2).

GAPDH used as a specificity and loading control.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-TDP43 antibody [2E2-D3] (ab57105)

Predicted band size: 44 kDa

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• sELISA

Supplier Data

Sandwich ELISA - Anti-TDP43 antibody [2E2-D3] (AB57105)

Detection limit for recombinant GST tagged TDP43 is 0.3 ng/ml as a capture antibody.

This image was generated using the ascites version of the product.

• WB

Supplier Data

Western blot - Anti-TDP43 antibody [2E2-D3] (AB57105)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-TDP43 antibody [2E2-D3] (ab57105) at 5 µg/mL

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with TDP43, cell lysate

Lane 2:

Untransfected HEK-293T cell lysate

Predicted band size: 44 kDa

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• WB

CiteAb

Western blot - Anti-TDP43 antibody [2E2-D3] (AB57105)

TDP43 western blot using anti-TDP43 antibody [2E2-D3] ab57105. Publication image and figure legend from Chandran, J. S., Sharp, P. S., et al., 2017, Sci Rep, PubMed 29116194.

ab57105 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab57105 please see the product overview.

Modulating HDAC4 expression enhances AAV9 transduction. (A) Western blot analysis of pulldowns between whole brain lysates and AAV9 (4 × 1011 vg for either virus) with streptavidin-agarose corroborates data from LC/MS. Note that endogenous SRSF1 expression was not detectable by conventional western blot, and so SRSF1-flag was overexpressed in HEK cells and incubated with the AAVs. Images shown are from multiple blots taken from a single pulldown probed for different interactors. Pulldowns were repeated at least three times for all targets shown. (B) Both AAV9 and AAV9-138 can bind to HDAC4 indicating that the HDAC4-AAV interaction does not arise purely from the capsid modification. Images shown are the same pulldown probed on different blots. (C) Two different miRNA sequences for HDAC4 demonstrate at least a 50% knockdown in HEK293T cells normalized to the control. (D) Western blot analysis of protein lysates taken 48 hrs after AAV-mcherry was added at 50,000 vg/cell show that the miHDAC4 #2 sequence augmented mcherry expression compared to a control miRNA scramble sequence. Tubulin immunoreactivity measured after stripping blot of mcherry signal. (E) Densitometry of western blots show that pretreating cells with the miHDAC4 #2 sequence yielded a 1.9 ± 0.48 (n = 5) fold increase in mcherry expression. (F) HDAC4 knockdown significantly (Student’s unpaired t-test used for each gene) reduces SF3B1, SRSF1, and SRSF5 levels in HEK293T cells stably expressing either a lentiviral miR-RNAi control or a miR-RNAi-HDAC4. (G,H) HDAC4 knockdown HEK cells show a significant increase in AAV9 particles in the nucleus compared to miR-RNAi controls at 20 minutes (Mann-Whitney test, n = 7, *p = 0.026) but not 60 minutes (Mann-Whitney test, miControl : n = 9; miHDAC4 : n = 5, p = 0.15). Corrected total cell fluorescence (CTCF) was used to quantify the AAV9-138 particles as described in methods. Arrows indicate regions where viral particles are clearly co-localizing with nucleolin. Error bars represent mean ± SEM. The AAV9-138 particles are stained red and nucleolin is stained in green. Scale bar 10 µm.

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