Anti-Nectin 2 antibody [EPR21124]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 2 antibody [EPR21124] (AB233384)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Nectin 2 with ab233384 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining in human testis (PMID : 23758976) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 2 antibody [EPR21124] (AB233384)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Nectin 2 with ab233384 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining in human breast cancer (PMID : 23758976) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin 2 antibody [EPR21124] (AB233384)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Nectin 2 with ab233384 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining in human liver (PMID : 23758976) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Nectin 2 antibody [EPR21124] (AB233384)

Western blot : Anti-Nectin 2 antibody [EPR21124] ab233384 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 64-75 kDa in Wild-type A549 UNBOILED cell lysates with no signal observed at this size in NECTIN2 knockout A549 UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Nectin 2 antibody [EPR21124] (ab233384) at 1/1000 dilution

Lane 1:

Wild-type A549 UNBOILED at 20 µg

Lane 2:

Western blot - Human NECTIN2 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-nectin2-knockout-a549-cell-line-ab288906'>ab288906</a>) at 20 µg

Lane 2:

NECTIN2 knockout A549 UNBOILED cell line

Lane 3:

MCF7 UNBOILED at 20 µg

Lane 4:

Raji UNBOILED at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 58 kDa

Observed band size: 64-75 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-Nectin 2 antibody [EPR21124] (AB233384)

Blocking/Dilution buffer : 5% NFDM/TBST.

Bands between 50- and 85-kDa are splice variants and the glycosylated form of nectin2, consistent with what has been described in the literature (PMID : 15108322 PMID : 12445825 PMID : 23750010 PMID : 12913096).

All lanes:

Western blot - Anti-Nectin 2 antibody [EPR21124] (ab233384) at 1/1000 dilution

Lane 1:

Human testis lysate at 20 µg

Lane 2:

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

Lane 3:

HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

A549 (human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

Human fetal kidney lysate at 10 µg

Secondary

Lanes 1 - 5:

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

Lane 6:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 58 kDa

Observed band size: 50-85 kDa

false

Exposure time: 3min