Anti-Notch1 抗体 [EP1238Y]

• WB

Lab

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627)

Western blot : Anti-NOTCH1 antibody [EP1238Y] (ab52627) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52627 was shown to bind specifically to NOTCH1. A band was observed at 110-120 kDa in wild-type HL-60 cell lysates with no signal observed at this size in NOTCH1 knockout cell line. To generate this image, wild-type and NOTCH1 knockout HL-60 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution

Lane 1:

Wild-type HL-60 cell lysate at 20 µg

Lane 2:

NOTCH1 knockout HL-60 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

NOTCH1 knockout HAP1 cell lysate at 20 µg

Lane 5:

MOLT-4 cell lysate at 20 µg

Lane 6:

HepG2 cell lysate at 20 µg

Observed band size: 110-120 kDa

true

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (AB52627)

Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using unpurified ab52627 at a dilution of 1/100.

An Alexa Fluor^® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor^® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (AB52627)

Immunohistochemical staining of paraffin-embedded human brain with unpurified ab52627 at a dilution of 1/100.

A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution

All lanes:

HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 272 kDa

Observed band size: 110-120 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (AB52627)

Immunohistochemical staining of paraffin-embedded human brain with purified ab52627 at a dilution of 1/150.

A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Notch1 antibody [EP1238Y] (AB52627)

Intracellular Flow Cytometry analysis of permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (2% PFA, pink) withpurified ab52627 at a 1/200 dilution, or negative control rabbit monoclonal IgG (green). The secondary antibody was FITC goat anti-rabbit.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (AB52627)

Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using purified ab52627 at a dilution of 1/150.

An Alexa Fluor^® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor^® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (AB52627)

Verification of gene expression array data by immunohistochemical analysis of Notch 1 expression in subcutaneous tumors and lung metastases from a human melanoma (MeWo) xenograft experiment in mice.

Immunohistochemical staining for Notch1 expression (ab52627, red) in subcutaneous tumors and lung metastases (both panels) of MeWo (Human malignant melanoma cell line) cells.

All scale bars : 50 μm.

• WB

Lab

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627)

Lanes 1 - 4 : Merged signal (red and green). Green - ab52627 observed at 110-120 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab52627 was shown to react with Notch1 in wild-type HAP1 cells in western blot. Loss of signal was observed when NOTCH1 knockout sample was used. Wild-type and NOTCH1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab52627 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1 µg/mL

Lane 1:

Wild-type HAP1 cell lysate at 40 µg

Lane 2:

NOTCH1 knockout HAP1 cell lysate at 40 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 272 kDa

Observed band size: 110-120 kDa

false

• WB

Supplier Data

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution

All lanes:

Mouse brain at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 272 kDa

Observed band size: 110-120 kDa

false

• WB

Lab

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627)

Lanes 1- 2 : Merged signal (red and green). Green - ab52627 observed at 110-120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NOTCH1 knockout HeLa cell line (<a href='/products/cell-lines/human-notch1-knockout-hela-cell-line-ab261762'>ab261762</a>)

Lane 2:

NOTCH1 knockout HeLa cell lysate at 20 µg

Predicted band size: 272 kDa

Observed band size: 110-120 kDa

false