Anti-Sumo 1 antibody [EP298]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical analysis of paraffin embedded Human Glioma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical analysis of paraffin embedded Human Urinary bladder transitional carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical staining of paraffin-embedded human thyroid carcinoma sections labelling Sumo 1 with purified ab133352 at dilution of 1 : 100. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical analysis of paraffin embedded normal Human tonsil tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical analysis of paraffin embedded Human Breast carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (AB133352)

Immunohistochemical analysis of paraffin-embedded Human papillary cell thyroid gland carcinoma tissue labelling Sumo 1 with un-purified ab133352 at 1/100 dilution.

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

• WB

Unknown

Western blot - Anti-Sumo 1 antibody [EP298] (AB133352)

All lanes:

Western blot - Anti-Sumo 1 antibody [EP298] (ab133352) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HL60 cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

NIH3T3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 11 kDa

false

• WB

Unknown

Western blot - Anti-Sumo 1 antibody [EP298] (AB133352)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-Sumo 1 antibody [EP298] (ab133352) at 1/10000 dilution

Lane 1:

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

Lane 2:

MCF-7 (human breast carcinoma) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryo) whole cell lysate at 20 µg

Lane 5:

C6 (rat glioma) whole cell lysate at 20 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 1/10000 dilution

Predicted band size: 11 kDa

Observed band size: 18-90 kDa

false

• WB

Lab

Western blot - Anti-Sumo 1 antibody [EP298] (AB133352)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Sumo 1 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : MCF-7 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab133352 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab133352 was shown to recognize Sumo 1 when Sumo 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Sumo 1 knockout samples were subjected to SDS-PAGE. ab133352 and ab8245 (loading control to GAPDH) were diluted 1/000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)

Predicted band size: 11 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Anti-Sumo 1 antibody [EP298] (AB133352)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Sumo 1 antibody [EP298] (ab133352) at 1/1000 dilution

Lane 1:

Western blot - Recombinant Human Sumo 1 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-1-protein-ab140417'>ab140417</a>) at 0.01 µg

Lane 2:

Western blot - Recombinant Human Sumo 2 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-2-protein-ab140420'>ab140420</a>) at 0.01 µg

Lane 3:

Recombinant Human Sumo 3 protein (<a href='/en-us/products/unavailable/recombinant-human-sumo-3-protein-ab140414'>ab140414</a>) at 0.01 µg

Lane 4:

Western blot - Recombinant Human Sumo 4 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-4-protein-ab157025'>ab157025</a>) at 0.1 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: 11 kDa

Observed band size: 16 kDa

true

Exposure time: 10s