Anti-Vitamin D Binding protein antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Vitamin D Binding protein antibody (AB65636)

ab65636 staining Vitamin D binding protein in U937 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab65636 at 5μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vitamin D Binding protein antibody (AB65636)

IHC image of Vitamin D Binding Protein staining in human kidney FFPE section, performed on a BondTM 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 ab65636, 1μg/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

• WB

Project

Western blot - Anti-Vitamin D Binding protein antibody (AB65636)

All lanes:

Western blot - Anti-Vitamin D Binding protein antibody (ab65636) at 1 µg/mL

Lane 1:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Lane 2:

Human ovary tissue lysate - total protein (<a href='/en-us/products/unavailable/human-ovary-tissue-lysate-total-protein-ab30222'>ab30222</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

Exposure time: 1min

• WB

Unknown

Western blot - Anti-Vitamin D Binding protein antibody (AB65636)

All lanes:

Western blot - Anti-Vitamin D Binding protein antibody (ab65636) at 1 µg/mL

All lanes:

Western blot - Native Human Vitamin D Binding protein (<a href='/en-us/products/proteins-peptides/native-human-vitamin-d-binding-protein-ab90920'>ab90920</a>) at 0.01 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: 53 kDa

true

Exposure time: 1min