Anti-BCKDHA antibody [EPR27003-11]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA antibody [EPR27003-11] (AB305168)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK293 tissue labeling BCKDHA with ab305168 at 1/5000 (0.264 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on (A) Wild-type HEK293T cell pellet; no staining on (B) BCKDHA knockout HEK293T (ab266439) cell pellet.The section was incubated with ab305168 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) was used.
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-BCKDHA antibody [EPR27003-11] (AB305168)

Immunohistochemical analysis of paraffin-embedded Human colorectal can tissue labeling BCKDHA with ab305168 at 1/2000 (0.264 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human colorectal cancer (PMID : 28501528). The section was incubated with ab305168 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) was used. 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-BCKDHA antibody [EPR27003-11] (AB305168)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling BCKDHA with ab305168 at 1/2000 (0.264 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on human liver (PMID : 3224821). The section was incubated with ab305168 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) was used. 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-BCKDHA antibody [EPR27003-11] (AB305168)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling BCKDHA with ab305168 at 1/2000 (0.264 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on human colon (PMID : 28501528). The section was incubated with ab305168 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) was used.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-BCKDHA antibody [EPR27003-11] (AB305168)

BCKDHA was immunoprecipitated from 0.35 mg HEK293T (human embryonic kidney epithelial cell), whole cell lysate 10 ug with ab305168 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305168 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HEK293T (human embryonic kidney epithelial cell), whole cell lysate 10 ug
Lane 2 : ab305168 IP in HEK293T whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305168 in HEK293T whole cell lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-BCKDHA antibody [EPR27003-11] (ab305168) at 1/1000 dilution

Lane 1:

HEK293T (human embryonic kidney epithelial cell), whole cell lysate 10 μg

Lane 2:

HEK293T 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: 49 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-BCKDHA antibody [EPR27003-11] (AB305168)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 26 seconds

All lanes:

Western blot - Anti-BCKDHA antibody [EPR27003-11] (ab305168) at 1/1000 dilution

All lanes:

Human liver tissue lysate 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 1/20000 dilution

Observed band size: 49 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-BCKDHA antibody [EPR27003-11] (AB305168)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTIn Western blot, ab305168 was shown to bind specifically to BCKDHA. A band was observed at 49 kDa in wild-type HEK293T cell lysates with no signal observed at this size in BCKDHA knockout cell line ab266439 (knockout cell lysate ab258324). Exposure time : 15 seconds

All lanes:

Western blot - Anti-BCKDHA antibody [EPR27003-11] (ab305168) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate 20 μg

Lane 2:

BCKDHA knockout HEK293T whole cell lysate 20 μg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 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 1/20000 dilution

Observed band size: 49 kDa

false

Exposure time: 15s