重组Anti-Hes1抗体[EPR4226]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (AB108937)

Primary ab concentration dilution : 1 : 200, (0.5ug/ml)

Secondary ab : ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG

Secondary ab concentration : Prediluted

Tissue : Human brain

Fixative : Paraffin-embedded sections

Counter stain : Hematoxylin

Antigen retrieval : Perform heat mediated antigen retrieval using Tris/EDTA Buffer, PH9

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (AB108937)

Immunohistochemical staining of Hes1 in paraffin-embedded human placenta tissue with unpurified ab108937 at 1/250 dilution.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (AB108937)

Immunohistochemical staining of paraffin embedded human brain with unpurified ab108937 at a working dilution of 1 in 90. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Hes1 antibody [EPR4226] (AB108937)

Overlay histogram showing SH-SY5Y cells stained with unpurifiedab108937 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab108937 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5000 events was performed.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hes1 antibody [EPR4226] (AB108937)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% TritonX-100 permeabilised SH-SY5Y (Human neuroblastoma epithelial cell) labelling Hes1 with ab108937 at 1/1000 dilution, followed by AlexaFluor^® 488 Goat anti-Rabbit secondary (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in SH-SY5Y cell line is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain alpha Tubulin at 1/200 dilution (red). The Nuclear counterstain was DAPI (blue).

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

• WB

Lab

Western blot - Anti-Hes1 antibody [EPR4226] (AB108937)

Blocking and diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Hes1 antibody [EPR4226] (ab108937) at 1/1000 dilution

Lane 1:

SH-SY5Y (Human bone marrow epithelial) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

Human brain tissue lysate at 20 µg

Lane 9:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/20000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Exposure time: 120s