Anti-Alpha-S1-casein antibody [EPR29093-719]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-S1-casein antibody [EPR29093-719] (AB323188)

Immunohistochemical analysis of paraffin-embedded Panel A 293T (human embryonic kidney epithelial cell) cells transfected with a Bovine Alpha-S1-casein expression vector containing a his tag. Panel B 293T cells transfected with empty vector containing a his tag. tissue labeling Alpha-S1-casein with ab323188 at 1/2000 (0.25ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) 293T transfected with a Bovine Alpha-S1-casein expression vector, no staining on (B) 293T transfected with empty vector.
The section was incubated with ab323188 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

• IP

Supplier Data

Immunoprecipitation - Anti-Alpha-S1-casein antibody [EPR29093-719] (AB323188)

Alpha-S1-casein was immunoprecipitated from 0.35 mg 293T cells transfected with a Bovine Alpha-S1-casein expression vector containing a myc-His-tag®, whole cell lysate with ab323188 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323188 at 1/1000 dilution.

Blocking and dilution buffer and concentration : Blocking Buffer diluted with an equal volume of TBS

Anti-Alpha-S1-casein antibody (ab323188) staining at 1/1000 dilution, shown in green(down); Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in magenta;IgG heavy chain show in green(up).

Avoid using milk-based buffers for blocking and dilution, as they might cause interference.

All lanes:

Immunoprecipitation - Anti-Alpha-S1-casein antibody [EPR29093-719] (ab323188) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab323188 at 1/30 IP in HEK-293 (human embryonic kidney epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323188 in 293T cellstransfected with a Bovine Alpha-S1-casein expression vectorcontaining a myc-His-tag®, whole cell 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

Observed band size: 34 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Alpha-S1-casein antibody [EPR29093-719] (AB323188)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.

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

Milk Powder Trim was diluted using 1%TBS buffer.

There is an empty lane between lane1 and lane2.

Western blot : Anti-Alpha-S1-casein antibody (ab323188) staining at 1/1000 dilution, shown in green.

Avoid using milk-based buffers for blocking and dilution, as they might cause interference.

All lanes:

Western blot - Anti-Alpha-S1-casein antibody [EPR29093-719] (ab323188) at 1/1000 dilution

Lane 1:

Milk whole lysate at 20 µg

Lane 2:

Milk whole lysate at 10 µg

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG H&L (800CW) at 1/100000 dilution

Lanes 1 - 2:

Goat Anti-Mouse IgG H&L (680RD) at 1/100000 dilution

Observed band size: 34 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Alpha-S1-casein antibody [EPR29093-719] (AB323188)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.

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

Western blot : Anti-Alpha-S1-casein antibody (ab323188) staining at 1/1000 dilution, shown in green(up); Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in magenta; Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution, shown in green(down).

Milk Powder Trim was diluted using 1%TBS buffer.

Avoid using milk-based buffers for blocking and dilution, as they might cause interference.

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

All lanes:

Western blot - Anti-Alpha-S1-casein antibody [EPR29093-719] (ab323188) 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 20 µg

Lane 2:

293T cells transfected with a Bovine Alpha-S1-casein expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG H&L (800CW) at 1/100000 dilution

Lanes 1 - 2:

Goat Anti-Mouse IgG H&L (680RD) at 1/100000 dilution

Observed band size: 34 kDa,36 kDa

false

Exposure time: 180s