Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free (AB315379)

This data was developed using ab315378, the same antibody clone in a different buffer formulation.

ASIC2 was immunoprecipitated from 0.35 mg Human cerebellum tissue lysate with ab315378 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315378 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-ASIC2 antibody [EPR25412-63] (<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a> at 1/30 IP in Human cerebellum tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a> in human cerebellum tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free (AB315379)

This data was developed using ab315378, the same antibody clone in a different buffer formulation.

Negative control : spleen, kidney, tonsil (PMID : 8631835, 10842183).

The observed MW is consistent with what has been described in the literature (PMID : 11842212, 37422581).

The identity of the lower MW band at approximately 50 kDa in Lane 4 is unknown.

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.

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

All lanes:

Western blot - Anti-ASIC2 antibody [EPR25412-63] (<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Lane 5:

Human hippocampus tissue lysate at 20 µg

Lane 6:

Human hypothalamus tissue lysate at 20 µg

Lane 7:

Human tonsil 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: 70 kDa,150 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free (AB315379)

This data was developed using ab315378, the same antibody clone in a different buffer formulation.

The observed MW is consistent with what has been described in the literature (PMID : 11842212 ).

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.

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

All lanes:

Western blot - Anti-ASIC2 antibody [EPR25412-63] (<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human spinal cord 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: 70 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free (AB315379)

This data was developed using ab315378, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with human ASIC1 and ASIC3.

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.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

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

All lanes:

Western blot - Anti-ASIC2 antibody [EPR25412-63] (<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human ASIC1 expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a human ASIC2 expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 4:

293T cells transfected with a human ASIC3 expression vector containing a myc-His-tag®, whole cell lysate at 10 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: 70 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-ASIC2 antibody [EPR25412-63] - BSA and Azide free (AB315379)

This data was developed using ab315378, the same antibody clone in a different buffer formulation.

Negative control : liver (PMID : 8631835, 10842183).

The observed MW is consistent with what has been described in the literature (PMID : 11842212 ).

The identity of the lower MW band at approximately 37 kDa in Lane 4 is unknown.

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.

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

All lanes:

Western blot - Anti-ASIC2 antibody [EPR25412-63] (<a href='/en-us/products/primary-antibodies/asic2-antibody-epr25412-63-ab315378'>ab315378</a>) at 1/1000 dilution

Lane 1:

Mouse spinal cord tissue lysate at 20 µg

Lane 2:

Mouse hippocampus tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Rat hippocampus tissue lysate at 20 µg

Lane 5:

Rat spinal cord 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: 70 kDa,36 kDa

false

Exposure time: 48s