Anti-DPP4 antibody [EPR20819]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DPP4 antibody [EPR20819] (AB215711)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD26 with ab215711 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on the bile canaliculi of human liver (PMID : 23361237) 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 before commencing with IHC staining protocol.

• I-ELISA

Lab

Indirect ELISA - Anti-DPP4 antibody [EPR20819] (AB215711)

ELISA analysis of Human DPP4 recombinant protein at 1000 ng/mL with ab215711. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-DPP4 antibody [EPR20819] (AB215711)

Blocking : 5% NFDM/TBST.

Exposure times :

Lanes 1,3 : 3 minutes

Lane 2 : 15 seconds

Lane 4 : 1 minute

Lane 5 : 30 seconds

The molecular weight observed is consistent with what has been described in the literature (PMID : 25589660).

All the lysates were preparaed using 1%SDS hot lysis method

All lanes:

Western blot - Anti-DPP4 antibody [EPR20819] (ab215711) at 1/1000 dilution

Lane 1:

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

Lane 2:

LoVo (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

Human fetal kidney at 10 µg

Lane 4:

Human fetal lung at 10 µg

Lane 5:

Human fetal liver at 10 µg

Secondary

Lane 1:

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 2:

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

Lanes 3 - 5:

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: 88 kDa

false

• WB

Lab

Western blot - Anti-DPP4 antibody [EPR20819] (AB215711)

Western blot : Rabbit Monoclonal[EPR20819] to DPP4 ab215711 staining at 1/200 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 110-130 kDa in Wild-type HCT 116 UNBOILED cell lysates with no signal observed at this size in DPP4 knockout HCT 116 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 BSA 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-DPP4 antibody [EPR20819] (ab215711) at 1/200 dilution

Lane 1:

Wild-type HCT 116 UNBOILED at 20 µg

Lane 2:

Western blot - Human DPP4 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-dpp4-knockout-hct116-cell-line-ab289063'>ab289063</a>) at 20 µg

Lane 2:

DPP4 knockout HCT 116 UNBOILED cell line at 20 µg

Lane 3:

Jurkat UNBOILED at 20 µg

Secondary

All lanes:

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

Predicted band size: 88 kDa

Observed band size: 110-130 kDa,50 kDa

false

• WB

Lab

Western blot - Anti-DPP4 antibody [EPR20819] (AB215711)

Blocking and diluting buffer : 5% NFDM/TBST.

For this product, we recommend 1%SDS hot lysis method.

For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

Exposure time :
Lane 1 : 3 minutes
Lane 2 : 7 seconds

All lanes:

Western blot - Anti-DPP4 antibody [EPR20819] (ab215711) at 0.4 µg/mL

Lane 1:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared using RIPA lysis method at 20 µg

Lane 2:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared using 1%SDS hot lysis method 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

Predicted band size: 88 kDa

false