Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue labelling TLS/FUS with ab283272 at 1/100 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain.

Nuclear staining on human lung carcinoma. The section was incubated with ab283272 for 30 mins at room temperature and followed mouse specific IgG antibody for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue labelling TLS/FUS with ab283272 at 1/100 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on human colon. The section was incubated with ab283272 for 30 mins at room temperature and followed mouse specific IgG antibody for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

ab283272 staining TLS/FUS in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes. The cells were then incubated with ab283272 at 1/50 dilution and ab179513 (Rabbit monoclonal to beta Tubulin) at 1/20 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150113 ) at 1/1000 dilution (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 1/500 dilution (shown in red). Nuclear DNA was labelled in blue with DAPI.

Confocal image showing nuclear staining in HeLa cell line.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue labelling TLS/FUS with ab283272 at 1/100 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain.

Nuclear staining on rat colon. The section was incubated with ab283272 for 30 mins at room temperature and followed mouse specific IgG antibody for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse stomach tissue labelling TLS/FUS with ab283272 at 1/100 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain.

Nuclear staining on mouse stomach. The section was incubated with ab283272 for 30 mins at room temperature and followed mouse specific IgG antibody for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

ab283272 staining TLS/FUS in NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes. The cells were then incubated with ab283272 at 1/50 dilution and ab179513 (Rabbit monoclonal to beta Tubulin) at 1/20 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150113 ) at 1/1000 dilution (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 1/500 dilution (shown in red). Nuclear DNA was labelled in blue with DAPI.

Confocal image showing nuclear staining in NIH/3T3 cell line.

• WB

Unknown

Western blot - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 30344044).

Lanes 1-2 : Merged signal (red and green). Green - ab283272 observed at 60 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab283272 Anti-FUS antibody was shown to react with FUS in HAP1 cells in Western blot. Loss of signal was observed when FUS knockout sample was used. Wild-type and FUS knockout samples were subjected to SDS-PAGE.

ab283272 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
70kDa band is the non-specific band according to KO validation.

All lanes:

Western blot - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (ab283272) at 1/1000 dilution

Lane 1:

Wild type HAP1(human chronic myelogenous leukemia near-haploid cell line), whole cell lysate at 20 µg

Lane 2:

FUS knockout HAP1(human chronic myelogenous leukemia near-haploid cell line), whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 60 kDa

false

• WB

Unknown

Western blot - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (AB283272)

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 30344044).
70kDa band is the non-specific band according to KO validation.

Lane 1 : 70 seconds
Lane 2 : 26 seconds

All lanes:

Western blot - Anti-TLS/FUS antibody [Z-FUS-5] - Mouse IgG2a (Chimeric) (ab283272) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 53 kDa

false