Anti-CLN5 antibody [EPR12197(B)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling CLN5 with purified ab170899 at 1/2000 dilution (0.22 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

Immunofluorescent analysis of A431 cells labeling CLN5 with unpurified ab170899 at 1/100 dilution (green). DAPI nuclear staining (blue).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

Immunocytochemistry/ Immunofluorescence analysis of T-47D (Human ductal breast epithelial tumor epithelial cell) cells labeling CLN5 with purified ab170899 at 1/50 dilution (8.7 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CLN5 with unpurified ab170899 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

Western blot : Rabbit Monoclonal[EPR12197(B)] to CLN5 ab170899 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 50 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in CLN5 knockout U-87 MG cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (ab170899) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cln5-antibody-epr12197b-bsa-and-azide-free-ab238931'>ab238931</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

CLN5 knockout U-87 MG at 20 µg

Lane 3:

HEK-293 at 20 µg

Lane 4:

Raji at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 45-60 kDa

false

• WB

Supplier Data

Western blot - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

All lanes:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (ab170899) at 1/1000 dilution

Lane 1:

Fetal kidney lysate at 10 µg

Lane 2:

T47-D cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 41 kDa

false

• WB

Unknown

Western blot - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

The molecular weights observed are consistent with what have been described in the literature (PMID : 26342652).

All lanes:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (ab170899) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 3:

A549 (Human lung carcinoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 48-52 kDa

false

• WB

Unknown

Western blot - Anti-CLN5 antibody [EPR12197(B)] (AB170899)

The molecular weights observed are consistent with what have been described in the literature (PMID : 26342652).

All lanes:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (ab170899) at 1/5000 dilution

All lanes:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 48-52 kDa

false