Anti-Prothrombin antibody [RM1252]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prothrombin antibody [RM1252] (AB322113)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Prothrombin with ab322113 at 1/5000 (0.243 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on plasma on human colon. The section was incubated with ab322113 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-Prothrombin antibody [RM1252] (AB322113)

Prothrombin was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab322113 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322113 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : ab322113 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab in HepG2 whole cell lysate

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17637839 and PMID : 21131592).

All lanes:

Immunoprecipitation - Anti-Prothrombin antibody [RM1252] (ab322113) at 1/30 dilution

All lanes:

HepG2 (human hepatocellular carcinoma epithelial 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: 84s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prothrombin antibody [RM1252] (AB322113)

Immunohistochemical analysis of paraffin-embedded (A) Rat non-perfused fixed kidney (B) Rat perfused fixed kidney tissue labeling Prothrombin with ab322113 at 1/2000 (0.243 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on plasma in (A) rat non-perfused fixed kidney and (B) rat perfused fixed kidney. The section was incubated with ab322113 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prothrombin antibody [RM1252] (AB322113)

Immunohistochemical analysis of paraffin-embedded (A) Mouse non-perfused fixed kidney (B) Mouse perfused fixed kidney tissue labeling Prothrombin with ab322113 at 1/2000 (0.243 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on plasma in (A) mouse non-perfused fixed kidney and (B) mouse perfused fixed kidney. The section was incubated with ab322113 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.

• WB

Supplier Data

Western blot - Anti-Prothrombin antibody [RM1252] (AB322113)

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

Negative control : HeLa (PMID : 1463733).

Low expression : cerebellum.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17637839 and PMID : 21131592).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Actual observed bands 33-72kDa.

All lanes:

Western blot - Anti-Prothrombin antibody [RM1252] (ab322113) at 1/1000 dilution

Lane 1:

Human plasma lysate at 4 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human cerebellum tissue lysate at 20 µg

Lane 4:

Human heart tissue lysate at 20 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

HeLa (human cervical 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

Observed band size: 33-72 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Prothrombin antibody [RM1252] (AB322113)

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

Low expression : Brain

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17637839 and PMID : 21131592).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Actual observed bands 33-72kDa.

Exposure times : left panel : 10 seconds; right panel : 147 seconds.

All lanes:

Western blot - Anti-Prothrombin antibody [RM1252] (ab322113) at 1/1000 dilution

Lane 1:

Mouse plasma lysate at 10 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Rat plasma lysate at 10 µg

Lane 7:

Rat liver tissue lysate at 20 µg

Lane 8:

Rat brain tissue lysate at 20 µg

Lane 9:

Rat heart 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 1/100000 dilution

Observed band size: 33-72 kDa,36 kDa

false