Anti-Nectin 3 antibody [EPR26326-22]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Nectin 3 with ab314219 at 1/500 (1.008 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Positive staining on human lung carcinoma. The section was incubated with ab314219 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling Nectin 3 with ab314219 at 1/500 (1.008 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Negative control : no staining on human cardiac muscle. The section was incubated with ab314219 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Nectin 3 with ab314219 at 1/500 (1.008 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Positive staining on human testis. The section was incubated with ab314219 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Nectin 3 with ab314219 at 1/500 (1.008 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Positive staining on human placenta. The section was incubated with ab314219 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

• IP

Supplier Data

Immunoprecipitation - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Nectin 3 was immunoprecipitated from 0.35 mg HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab314219 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314219 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate Lane 2 : ab314219 IP in HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314219 in HCT116 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 137 seconds The identity of the lower MW band at approximately 25 kDa (in lane 2) is unknown.

All lanes:

Immunoprecipitation - Anti-Nectin 3 antibody [EPR26326-22] (ab314219) at 1/30 dilution

All lanes:

HCT116 (human colorectal 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: 137s

• WB

Supplier Data

Western blot - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : U-937 (PMID : 11024295). Nectin 3 is a glycoprotein of approximately 100 kDa and detected as a 60 kDa band after treated with Protein Deglycosylation MIX II. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lane 1-2 : 10 seconds, Lane 3-5 : 5 seconds.

All lanes:

Western blot - Anti-Nectin 3 antibody [EPR26326-22] (ab314219) at 1/1000 dilution

Lane 1:

Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HCT 116 whole cell lysate treated with Protein Deglycosylation MIX II at 15 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 5:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 60-100 kDa

false

• WB

Supplier Data

Western blot - Anti-Nectin 3 antibody [EPR26326-22] (AB314219)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : heart (PMID : 11024295), colon (PMID : 11024295). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-Nectin 3 antibody [EPR26326-22] (ab314219) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human colon tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 60-100 kDa

false

Exposure time: 10s