Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (AB238931)

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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170899).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (AB238931)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (unpurified ab170899).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (AB238931)

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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170899).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (AB238931)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (unpurified ab238931).

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)] - BSA and Azide free (AB238931)

This data was developed using ab170899, the same antibody clone in a different buffer formulation.

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)] (<a href='/en-us/products/primary-antibodies/cln5-antibody-epr12197b-ab170899'>ab170899</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (ab238931) 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

Unknown

Western blot - Anti-CLN5 antibody [EPR12197(B)] - BSA and Azide free (AB238931)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170899).

All lanes:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (<a href='/en-us/products/primary-antibodies/cln5-antibody-epr12197b-ab170899'>ab170899</a>) 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)] - BSA and Azide free (AB238931)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170899).

All lanes:

Western blot - Anti-CLN5 antibody [EPR12197(B)] (<a href='/en-us/products/primary-antibodies/cln5-antibody-epr12197b-ab170899'>ab170899</a>) 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