Anti-KLC1 antibody [EPR12441(B)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling KLC1 with purified ab174273 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Immunofluorescent analysis of U87-MG cells labeling KLC1 with ab174273 at 1/50 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Intracellular flow cytometric analysis of permeabilized HeLa cells labeling KLC1with ab174273 at 1/10 dilution (red) or a rabbit IgG (negative) (green).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling KLC1 with ab174273 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Western blot analysis on immunoprecipitation pellet from SH-SY5Y cell lysate using ab174273 at 1/10 dilution.

All lanes:

Immunoprecipitation - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

Predicted band size: 65 kDa

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• WB

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Western blot - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

Lanes 1-3 : Merged signal (red and green). Green - ab174273 observed at 65 kDa. Red - loading control ab8245 observed at 36 kDa.

ab174273 Anti-KLC1 antibody [EPR12441(B)] was shown to specifically react with KLC1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265284 (knockout cell lysate ab257496) was used. Wild-type and KLC1 knockout samples were subjected to SDS-PAGE. ab174273 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

KLC1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human KLC1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-klc1-knockout-hela-cell-line-ab265284'>ab265284</a>)

Lane 3:

U-87 MG cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

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• WB

Supplier Data

Western blot - Anti-KLC1 antibody [EPR12441(B)] (AB174273)

All lanes:

Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273) at 1/1000 dilution

Lane 1:

U87-MG cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Predicted band size: 65 kDa

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