PA5-78764

antibody from Invitrogen Antibodies

Targeting: ALPP PALP, PLAP

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-78764

Provider

Invitrogen Antibodies

Product name

Placental Alkaline Phosphatase Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Alkaline Phosphatase in human placenta extract (lane 1). Sample was incubated with Alkaline Phosphatase polyclonal antibody (Product # PA5-78764) at a dilution of 0.5 µg/mL. Signal development was performed using a chemiluminescence (ECL) kit.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ALPP in, Lane 1: human placenta tissue lysates, Lane 2: human placenta tissue lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with Placental Alkaline Phosphatase Polyclonal Antibody (Product # PA5-78764) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for ALPP at approximately 70 kDa. The expected band size for ALPP is at 58 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of ALPP using anti- ALPP antibody (Product # PA5-78764) . ALPP was detected in a section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 5μg/mL rabbit anti- ALPP antibody (Product # PA5-78764) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Alkaline Phosphatase on paraffin-embedded human placenta tissue. Sample was incubated with Alkaline Phosphatase polyclonal antibody (Product# PA5-78764) with a dilution of 1 µg/mL, and developed by Streptavidin-Biotin-Complex (SABC) method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of ALPP in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/mL rabbit anti-ALPP antibody (Product # PA5-78764) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of Placental Alkaline Phosphatase in SiHa cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with Placental Alkaline Phosphatase Polyclonal Antibody (Product # PA5-78764) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.