重组Anti-UFM1抗体[EPR4264(2)]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

ab109305 staining UFM1 in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

ab109305 staining UFM1 in human thyroid carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab109305 at 1/250 dilution.

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

• IP

Lab

Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

ab109305 immunoprecipitating UFM1. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

Lane 1 : 293 (Human embryonic kidney epithelial cell) whole cell lysate 10ug
Lane 2 : 293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab109305 in 293 (Human embryonic kidney epithelial cell) whole cell lysate

All lanes:

Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

Predicted band size: 9 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

ab109305 staining UFM1 in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

ab109305 staining UFM1 in mouse mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• WB

Lab

Western blot - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

Western blot : Anti-UFM1 antibody [EPR4264(2)] (ab109305) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab109305 was shown to bind specifically to UFM1. A band was observed at 9/28 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in UFM1 knockout cell line. To generate this image, wild-type and UFM1 knockout HEK-293T 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

UFM1 knockout HEK-293T cell lysate at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 9 kDa,28 kDa

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• WB

Unknown

Western blot - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

All lanes:

Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/1000 dilution

Lane 1:

Fetal kidney lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Predicted band size: 9 kDa

Observed band size: 9 kDa

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• WB

Lab

Western blot - Anti-UFM1 antibody [EPR4264(2)] (AB109305)

Mouse heart cell lysate Blocking and diluting buffer : 5% NFDM/TBST

The band at around 28kDa might be UFM1 conjugates [PMID : 23152784].

All lanes:

Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/2000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Mouse spleen cell lysate at 20 µg

Lane 4:

Mouse heart cell lysate at 20 µg

Lane 5:

Rat spleen cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 9 kDa

Observed band size: 28 kDa,9 kDa

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