Anti-EGFL7 antibody [EPR22603-113]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFL7 antibody [EPR22603-113] (AB256451)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling EGFL7 with ab256451 at 1/500 dilution (1.12 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human placenta (PMID : 24751645) is observed. The section was incubated with ab256451 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-EGFL7 antibody [EPR22603-113] (AB256451)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (Human umbilical vein endothelial cell) cells labeling EGFL7 with ab256451 at 1/50, (10 μg/ml), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cells. Negative control : Jurkat (PMID : 15162510). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFL7 antibody [EPR22603-113] (AB256451)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling EGFL7 with ab256451 at 1/500 dilution (1.12 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on blood vessels in human pancreas (PMID : 18497746) is observed. The section was incubated with ab256451 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• WB

Lab

Western blot - Anti-EGFL7 antibody [EPR22603-113] (AB256451)

Western blot : Anti-EGFL7 antibody [EPR22603-113] ab256451 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 30 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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-EGFL7 antibody [EPR22603-113] (ab256451) at 1/1000 dilution

Lane 1:

Wild-type A549 BFA (5g/mL,6h) at 20 µg

Lane 2:

Wild-type A549 BFA (0 g/mL,6h) at 20 µg

Lane 3:

EGFL7 knockout A549 BFA (5 g/mL, 6 h) at 20 µg

Lane 4:

EGFL7 knockout A549 BFA (0 g/mL, 6 h) at 20 µg

Lane 5:

HUVEC at 20 µg

Lane 6:

Jurkat at 20 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• WB

Lab

Western blot - Anti-EGFL7 antibody [EPR22603-113] (AB256451)

The molecular weight observed is consistent with what has been described in the literature (PMID : 18497746).

Negative control : Jurkat (PMID : 15162510).

Exposure time : 3 minutes.

Blocking and Diluting Buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-EGFL7 antibody [EPR22603-113] (ab256451) at 1/1000 dilution

Lane 1:

Human placenta lysate at 20 µg

Lane 2:

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

Lane 3:

Jurkat (human t cell leukemia t lymphocyte) whole cell 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 1/20000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false