Anti-ITM2B antibody [EPR27849-47]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ITM2B antibody [EPR27849-47] (AB315234)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells labelling ITM2B with ab315234 at 1/50 dilution (1 ug, Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ITM2B antibody [EPR27849-47] (AB315234)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling ITM2B with ab315234 at 1/50 dilution (1 ug, Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-ITM2B antibody [EPR27849-47] (AB315234)

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

Low expression : liver, skeletal muscle (PMID : 10391242)

55 kDa pro-ITM2B and 42-17 kDa ITM2B band were observed. Bands beneath 42 kDa were cleavage fragments (PMID : 24524963).

Samples are non-boiled as boiling may cause protein aggregation.

There is an unknown band at 120 kDa.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ITM2B antibody [EPR27849-47] (ab315234) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Lane 5:

SH-SY5Y (human neuroblastoma epithelial cell) 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 1/100000 dilution

Observed band size: 55 kDa,42-17 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ITM2B antibody [EPR27849-47] (AB315234)

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

Low expression : skeletal muscle (PMID : 12566161, PMID : 10391242)

55 kDa pro-ITM2B and 42-17 kDa ITM2B band were observed. Bands beneath 42 kDa were cleavage fragments (PMID : 24524963).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ITM2B antibody [EPR27849-47] (ab315234) at 1/1000 dilution

Lane 1:

Rat hippocampus tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle tissue lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/100000 dilution

Observed band size: 55 kDa,42-17 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ITM2B antibody [EPR27849-47] (AB315234)

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

Low expression : liver, spleen, testis (PMID : 12566161).

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 24524963, PMID : 28131015)

55 kDa pro-ITM2B and 42-17 kDa ITM2B band were observed. Bands beneath 42 kDa were cleavage fragments (PMID : 24524963).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ITM2B antibody [EPR27849-47] (ab315234) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat spleen tissue 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 1/100000 dilution

Observed band size: 55 kDa,42-17 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-ITM2B antibody [EPR27849-47] (AB315234)

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

Low expression : spleen, testis, liver (PMID : 12566161, PMID : 10391242).

55 kDa pro-ITM2B and 42-17 kDa ITM2B band were observed. Bands beneath 42 kDa were cleavage fragment (PMID : 24524963).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ITM2B antibody [EPR27849-47] (ab315234) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Rat hippocampus tissue lysate at 20 µg

Lane 6:

Rat testis tissue 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 1/100000 dilution

Observed band size: 55 kDa,42-17 kDa,36 kDa

false

Exposure time: 81s