Anti-Ihh antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Ihh antibody (AB39634)

ICC/IF image of ab39634 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab39634, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ihh antibody (AB39634)

IHC image of Ihh staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab39634, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• IHC-P

AbReview11757****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ihh antibody (AB39634)

ab39634 (1/50) staining Ihh in paraffin-embbeded mouse pancreas tissue sections. Tissue underwent fixation in formaldehyde, heat-mediated antigen retrieval in citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block and 10 minutes/protein block). For further experimental details please refer to abreview. Strongest staining observed in islet cells of the pancreas.

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.

• WB

Project

Western blot - Anti-Ihh antibody (AB39634)

The Ihh protein has a predicted molecular weight of 45 kDa. The first 27 amino acids of the Ihh sequence act as a signal sequence, and when cleaved the protein has an expected molecular weight of 42 kDa.

All lanes:

Western blot - Anti-Ihh antibody (ab39634) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Western blot - Mouse colon tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-colon-tissue-lysate-total-protein-ab29544'>ab29544</a>) at 10 µg

Lane 3:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 42 kDa

false

• WB

Project

Western blot - Anti-Ihh antibody (AB39634)

All lanes:

Western blot - Anti-Ihh antibody (ab39634) at 1 µg/mL

All lanes:

Lung (Human) Whole Cell Lysate - fetal normal tissue (<a href='/en-us/products/unavailable/lung-human-whole-cell-lysate-fetal-normal-tissue-ab30282'>ab30282</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

true

Exposure time: 30s