Anti-Annexin-7/ANXA7 antibody [EPR17090]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling ANXA7 with ab197586 at 1/100 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green). Cytoplasm staining on SH-SY5Y cell line is observed. Subcellular location is citied as Cytoplasm [J Cell Sci. 1995 Jan;108 ( Pt 1) : 85-95.] The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).

The negative controls are as follows : -
-ve control 1 - ab179586 at 1/100 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ANXA7 with ab197586 at 1/600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Cytoplasm staining on Human cervix carcinoma tissue is observed. Subcellular location citied as Cytoplasm [J Cell Sci. 1995 Jan;108 ( Pt 1) : 85-95.]. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary ab secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma from bone marrow) cellslabeling ANXA7 with ab197586 at 1/60 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ANXA7 with ab197586 at 1/600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Cytoplasm staining on Mouse liver tissue is observed. Subcellular location citied as Cytoplasm [J Cell Sci. 1995 Jan;108 ( Pt 1) : 85-95.]. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary ab secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cells) cells labeling ANXA7 with ab197586 at 1/100 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green). Cytoplasm staining on Neuro-2a cell line is observed. Subcellular location is citied as Cytoplasm [J Cell Sci. 1995 Jan;108 ( Pt 1) : 85-95.] The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).

The negative controls are as follows : -
-ve control 1 - ab179586 at 1/100 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

• WB

Supplier Data

Western blot - Anti-Annexin-7/ANXA7 antibody [EPR17090] (AB197586)

Blocking and diluting buffer was 5% NFDM /TBST

All lanes:

Western blot - Anti-Annexin-7/ANXA7 antibody [EPR17090] (ab197586) at 1/2000 dilution

Lane 1:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 10 µg

Lane 2:

SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate at 10 µg

Lane 3:

Rat brain lysate at 10 µg

Lane 4:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 5:

Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 6:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

Exposure time: 3min