Anti-Islet 1 antibody [EPR10362] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas tissue sections labeling Islet 1 with purified ab178400 at 1/6000 dilution (0.02 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Secondary only control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human small cell lung carcinoma tissue labeling Islet 1 with ab178400 at a 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human gastric neuroendocrine carcinoma tissue labeling Islet 1 with ab178400 at a 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

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

• IP

Supplier Data

Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

Western blot analysis on Immunoprecipitation pellet from either 1) SH-SY5Y cell lysate, or 2) 1xPBS (negative control); showing Islet 1, using ab178400 at 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

All lanes:

Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] - Neural Stem Cell Marker (<a href='/en-us/products/primary-antibodies/islet-1-antibody-epr10362-neural-stem-cell-marker-ab178400'>ab178400</a>)

Predicted band size: 39 kDa

true

• IP

Unknown

Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

ab178400 (purified ) at 1/20 dilution (1ug) immunoprecipitating Islet 1 in SH-SY5Y whole cell lysate.

Lane 1 (input) : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab178400 & SH-SY5Y whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab178400 in SH-SY5Y whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

All lanes:

Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] - Neural Stem Cell Marker (<a href='/en-us/products/primary-antibodies/islet-1-antibody-epr10362-neural-stem-cell-marker-ab178400'>ab178400</a>)

Predicted band size: 39 kDa

false

• WB

Supplier Data

Western blot - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (AB238919)

Lanes 1 - 4 : Merged signal (red and green). Green - ab178400 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab178400 was shown to recognize Islet 1 in wild-type HAP1 cells as signal was lost at the expected MW in Islet 1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Islet 1 knockout samples were subjected to SDS-PAGE. ab178400 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

Lanes 1 - 3:

Western blot - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (ab238919) at 1/1000 dilution

Lane 4:

Western blot - Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (ab238919) at 1000 µg

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

Islet 1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

SH-SY5Y whole cell lysate at 20 µg

Lane 4:

HepG2 whole cell lysate at 20 µg

Predicted band size: 39 kDa

false