All lanes : Anti-ZPI antibody [EPR23683-118] (ab273627) at 1/1000 dilution

Lane 1 : Human serum
Lane 2 : Human plasma

Lysates/proteins at 20 µl per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

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

The expression profile observed is consistent with what has been described in the literature (PMID:10460162, 9689066).

Exposure time: 3 minutes.

All lanes : Anti-ZPI antibody [EPR23683-118] (ab273627) at 1/1000 dilution

Lane 1 : Human liver cancer tissue lysate
Lane 2 : Human brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

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

The expression profile observed is consistent with what has been described in the literature (PMID: 10460162, 9689066). 

Negative control: Human brain (PMID: 10460162). 

This blot was developed using a higher sensitivity ECL substrate.

 Exposure time: 37 seconds.

All lanes : Anti-ZPI antibody [EPR23683-118] (ab273627) at 1/1000 dilution

Lane 1 : Mouse plasma
Lane 2 : Rat serum
Lane 3 : Rat plasma

Lysates/proteins at 20 µl per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 50 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

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

The expression profile observed is consistent with what has been described in the literature (PMID:10460162, 9689066).

Exposure times: Lane 1: 26 seconds; Lane 2-3: 5.5 seconds.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling ZPI with ab273627 at 1/1000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human liver (PMID: 20024489). The section was incubated with ab273627 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling ZPI with ab273627 at 1/1000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on pancreas islets (PMID: 16728581). The section was incubated with ab273627 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling ZPI with ab273627 at 1/1000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on plasma in human placenta. The section was incubated with ab273627 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling ZPI with ab273627 at 1/1000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat liver (PMID: 20024489). The section was incubated with ab273627 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling ZPI with ab273627 at 1/1000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The section was incubated with ab273627 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control: No staining on human cererbum (PMID: 20024489). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

ZPI was immunoprecipitated from 0.35 mg human plasma with ab273627 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273627 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Human plasma lysate 10µg

Lane 2: ab273627 IP in human plasma 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab273627 in human plasma 

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

Exposure time: 3 minutes.

The expression profile observed is consistent with what has been described in the literature (PMID:10460162).