Anti-MICU2 抗体 [EPR28910-9] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling MICU2 with ab320644 at 1/500 (0.998 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression tissue : No staining in human liver (PMID : 33114428). The section was incubated with ab320644 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling MICU2 with ab320644 at 1/500 (0.998 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human breast cancer. The section was incubated with ab320644 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling MICU2 with ab320644 at 1/500 (0.998 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human gastric cancer. The section was incubated with ab320644 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MICU2 with ab320644 at 1/500 (0.998 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human colon. The section was incubated with ab320644 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The identity of the higher MW bands at approximately 250 kDa are unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MICU2 antibody [EPR28910-9] (<a href='/products/primary-antibodies/micu2-antibody-epr28910-9-ab320644'>ab320644</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting MICU whole 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

Observed band size: 40 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Western blot : Anti-MICU2 antibody [EPR28910-9] ab320644 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 40 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in MICU2 knockout HCT 116 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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.

All lanes:

Western blot - Anti-MICU2 antibody [EPR28910-9] (<a href='/products/primary-antibodies/micu2-antibody-epr28910-9-ab320644'>ab320644</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

MICU2 knockout HCT 116 at 20 µg

Lane 3:

SW 480 at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 40 kDa

false

• WB

Supplier Data

Western blot - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : liver (PMID : 33114428) The identity of the higher MW bands above 75 kDa are unknown. The identity of the lower MW band at approximately 15 kDa is unknown. Lanes 3-5 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution. Exposure time : Lanes 1-2 : 180 seconds; Lanes 3-5 : 37 seconds.

All lanes:

Western blot - Anti-MICU2 antibody [EPR28910-9] (<a href='/products/primary-antibodies/micu2-antibody-epr28910-9-ab320644'>ab320644</a>) at 1/1000 dilution

Lane 1:

Human liver cancer tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Mouse liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 3 - 5:

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

Observed band size: 40 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30242232, PMID : 30504268). The identity of the higher MW bands above 75 kDa are unknown. The identity of the lower MW band at approximately 15 kDa is unknown. Exposure time : Lanes 1-3 : 37 seconds; Lanes 4-5 : 136 seconds.

All lanes:

Western blot - Anti-MICU2 antibody [EPR28910-9] (<a href='/products/primary-antibodies/micu2-antibody-epr28910-9-ab320644'>ab320644</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) whole cell lysate at 20 µg

Lane 5:

4T1 (mouse mammary gland carcinoma epithelial cell) whole 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

Observed band size: 40 kDa

false

• WB

Supplier Data

Western blot - Anti-MICU2 antibody [EPR28910-9] - BSA and Azide free (AB320645)

This data was developed using ab320644, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody doesn't cross-react with human MICU3. In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade  (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-MICU2 antibody [EPR28910-9] (<a href='/products/primary-antibodies/micu2-antibody-epr28910-9-ab320644'>ab320644</a>) at 1/1000 dilution

Lane 1:

His tagged human MICU2 protein at 5 ng

Lane 2:

His tagged human MICU3 protein at 50 ng

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

Observed band size: 40 kDa

false

Exposure time: 15s