Anti-ASAH1 antibody [EPR24476-30]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on bladder cancer. The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human colon tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon (PMID : 29920241). The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer. The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon cancer. The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling ASAH1 with ab282276 at 1/5000 dilution (0.118 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab282276 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

ASAH1 was immunoprecipitated from 0.35 mg SK-MEL-28 (human malignant melanoma cell ) whole cell lysate with ab322109 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322109 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SK-MEL-28 (human malignant melanoma cell ) whole cell lysate
Lane 2 : ab322109 IP in SK-MEL-28 (human malignant melanoma cell ) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322109 in SK-MEL-28 whole cell lysate

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

The IP experiment was performed by ab322109 using SK-MEL-28 lysate. The blot was probed by another anti-ASAH1 antibody (ab282276)(1 : 1000 dilution).

All lanes:

Immunoprecipitation - Anti-ASAH1 antibody [MJF-D29767-517] (<a href='/en-us/products/primary-antibodies/asah1-antibody-mjf-d29767-517-ab322109'>ab322109</a>) at 1/30 dilution

All lanes:

SK-MEL-28 (human malignant melanoma cell ) 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

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 7744740, 26553872, 30254208).

Exposure time : 70 seconds

All lanes:

Western blot - Anti-ASAH1 antibody [EPR24476-30] (ab282276) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 40 µg

Lane 2:

Human spleen tissue lysate at 40 µg

Lane 3:

Human heart tissue lysate at 40 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/2000 dilution

Predicted band size: 45 kDa

Observed band size: 36 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 7744740, 26553872, 30254208).

Negative control : A375 (PMID : 26553872, 30254208); MDA-MB-231 (PMID : 26553872).

Exposure time : 37 seconds

All lanes:

Western blot - Anti-ASAH1 antibody [EPR24476-30] (ab282276) at 1/1000 dilution

Lane 1:

SK-MEL-28 (human malignant melanoma) whole cell lysate at 20 µg

Lane 2:

MeWo (human malignant melanoma fibroblast) whole cell lysate at 20 µg

Lane 3:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - 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

Predicted band size: 45 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Anti-ASAH1 antibody [EPR24476-30] (AB282276)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 7744740, 26553872, 30254208).

Exposure time : 147 seconds

All lanes:

Western blot - Anti-ASAH1 antibody [EPR24476-30] (ab282276) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 40 µg

Lane 2:

Mouse colon tissue lysate at 40 µg

Lane 3:

Rat spleen tissue lysate at 40 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

Predicted band size: 45 kDa

Observed band size: 36 kDa

false