Anti-CSL4 antibody [EPR13526] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cervix tissue labeling CSL4 with ab181108 at 1/100 dilution. Counter stain Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CSL4 with ab181108 at 1/100 dilution. Counter stain Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde fixed 293 cells labeling CSL4 with ab181108 at 1/250 dilution. Secondary antibody Goat anti rabbit IgG (Alexa Fluor®488), Conterstained with DAPI blue.

• IP

Supplier Data

Immunoprecipitation - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108&44; the same antibody clone in a different buffer formulation.

Western blot analysis of CSL4 in immunoprecipitation pellets from Raji lysate. ab181108 used at 1/60 dilution. Secondary antibody Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500. Lane 2 shows negative control.

All lanes:

Immunoprecipitation - Anti-CSL4 antibody [EPR13526] (<a href='/en-us/products/primary-antibodies/csl4-antibody-epr13526-ab181108'>ab181108</a>)

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CSL4 antibody [EPR13526] (<a href='/en-us/products/primary-antibodies/csl4-antibody-epr13526-ab181108'>ab181108</a>) at 1/2000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

C6 cell lysate at 10 µg

Lane 3:

Raw264.7 cell lysate at 10 µg

Lane 4:

PC-12 cell lysate at 10 µg

Lane 5:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

False colour image of Western blot : Anti-CSL4 antibody [EPR13526] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab181108 was shown to bind specifically to CSL4. A band was observed at 22 kDa in wild-type HeLa cell lysates with no signal observed at this size in EXOSC1 CRISPR-Cas9 edited cell line ab265194 (EXOSC1 CRISPR-Cas9 edited cell lysate ab257945). The band observed in the CRISPR-Cas9 edited lysate lane above 22 kDa is likely to represent CSL4 with an insertion. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and EXOSC1 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CSL4 antibody [EPR13526] (<a href='/en-us/products/primary-antibodies/csl4-antibody-epr13526-ab181108'>ab181108</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

EXOSC1 CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 3:

PC-12 cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 21 kDa

Observed band size: 22 kDa

false

• WB

Supplier Data

Western blot - Anti-CSL4 antibody [EPR13526] - BSA and Azide free (AB250355)

This data was developed using ab181108, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CSL4 antibody [EPR13526] (<a href='/en-us/products/primary-antibodies/csl4-antibody-epr13526-ab181108'>ab181108</a>) at 1/10000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

293 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false