人RHOT2 (MIRO2) knockout HeLa cell裂解物

• Sanger seq

Unknown

Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell lysate (AB257639)

Allele-2 : 1 bp deletion in exon2

• Sanger seq

Unknown

Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell lysate (AB257639)

Allele-1 : 2 bp deletion in exon2

• WB

Supplier Data

Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (AB257639)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The samples were run on a Bis-Tris gel under reducing conditions.

Western blot : Anti-MIRO2 antibody (ab322962) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in magenta.

In Western blot, ab322962 was shown to bind specifically to MIRO2. Target of interest was observed at 80 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in MIRO2 knockout cell line (lane 2, knockout cell line ab265801 / knockout cell lysate ab257639). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

All lanes:

Western blot - Anti-MIRO2 antibody [RM2077] (<a href='/products/primary-antibodies/miro2-antibody-rm2077-ab322962'>ab322962</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639) at 20 µg

Lane 3:

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

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole 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: 80 kDa

false

• WB

Supplier Data

Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (AB257639)

Blocking buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS. Diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-MIRO2 antibody (ab320739) 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, ab320739 was shown to bind specifically to MIRO2. Target of interest was observed at 80 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size MIRO2 knockout cell line (lane 2, knockout cell line ab265801 / knockout cell lysate ab257639). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane.  Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C.  Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged. Exposure time : N/A

All lanes:

Western blot - Anti-MIRO2 antibody [EPR29118-85] (<a href='/products/primary-antibodies/miro2-antibody-epr29118-85-ab320739'>ab320739</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639) at 20 µg

Lane 3:

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

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole 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: 80 kDa

false