Anti-Cystatin D antibody [EPR25935-47] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Cystatin D antibody [EPR25935-47] - BSA and Azide free (AB305063)

This data was developed using ab305062, the same antibody clone in a different buffer formulation. Cystatin D was immunoprecipitated from 0.35 mg Human submandibular gland tissue lysate 10 µg with ab305062 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305062 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Human submandibular gland tissue lysate 10 µg Lane 2 : ab305062 IP in Human submandibular gland tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305062 in human submandibular gland tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds The identity of the higher MW band at approximately 75 kDa (in lane 1) is unknown.

All lanes:

Immunoprecipitation - Anti-Cystatin D antibody [EPR25935-47] (<a href='/en-us/products/primary-antibodies/cystatin-d-antibody-epr25935-47-ab305062'>ab305062</a>) at 1/1000 dilution

Lane 1:

Human submandibular gland tissue lysate 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/cystatin-d-antibody-epr25935-47-ab305062'>ab305062</a> IP in Human submandibular gland 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

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Cystatin D antibody [EPR25935-47] - BSA and Azide free (AB305063)

This data was developed using ab305062, the same antibody clone in a different buffer formulation.

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

Negative control : colon (PMID : 11879580).

The identity of the higher MW band at approximately 75 kDa is unknown.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Cystatin D antibody [EPR25935-47] (<a href='/en-us/products/primary-antibodies/cystatin-d-antibody-epr25935-47-ab305062'>ab305062</a>) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human submandibar gland 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 1/20000 dilution

Observed band size: 14 kDa

true

Exposure time: 114s

• WB

Supplier Data

Western blot - Anti-Cystatin D antibody [EPR25935-47] - BSA and Azide free (AB305063)

This data was developed using ab305062, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 180 seconds

All lanes:

Western blot - Anti-Cystatin D antibody [EPR25935-47] (<a href='/en-us/products/primary-antibodies/cystatin-d-antibody-epr25935-47-ab305062'>ab305062</a>) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with an empty vector containing a his tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human Cystatin D expression vector containing a his tag, 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: 17 kDa

false

Exposure time: 180s