PA5-47419

antibody from Invitrogen Antibodies

Targeting: ALPL HOPS, TNALP, TNAP, TNSALP

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-47419

Provider

Invitrogen Antibodies

Product name

ALPL Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs and Western blots, approximately 60% cross-reactivity with recombinant human ALPL is observed. Reconstitute at 0.2 mg/mL in sterile PBS.

Reactivity

Human, Mouse

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.2 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis from lysates of D3 mouse embryonic stem cell line, MEF mouse embryonic feeder cells, and mouse embryo tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-mouse Alkaline Phosphatase/ALPL Antigen Affinity-purified Polyclonal Antibody (Product # PA5-47419) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody. Specific bands were detected for Alkaline Phosphatase/ALPL at approximately 75-80 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ALPL in D3 mouse embryonic stem cell line, MEF mouse embryonic feeder cells, and mouse embryo tissue. Samples were incubated in ALPL polyclonal antibody (Product # PA5-47419) using a dilution of 1 µg/mL followed by a HRP-conjugated Anti-Goat IgG secondary antibody. Specific bands were detected for Alkaline Phosphatase/ALPL at approximately 75-80 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

KD of ALPL was achieved by transfecting U-2 OS with ALPL specific siRNAs (Silencer® select Product # s1301, s1300). Western blot analysis (Fig. a) was performed using whole cell extracts from the ALPL KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with ALPL Polyclonal Antibody (Product # PA5-47419, 1µg/mL) and Rabbit Anti-Goat IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27014, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to ALPL..

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ALPL Polyclonal Antibody (Product # PA5-47419) and a 57 kDa band corresponding to ALPL was observed across the cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), U-2 OS (Lane 2), SH-SY5Y (Lane 3) and MOLT-4 (Lane 4) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1µg/mL) and detected by chemiluminescence Rabbit Anti-Goat IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of ALPL in immersion fixed rat mesenchymal stem cells. Samples were incubated in ALPL polyclonal antibody (Product # PA5-47419) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of ALPL in Rat Mesenchymal Stem Cells. Samples were incubated in ALPL polyclonal antibody (Product # PA5-47419) or isotype control antibody followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody.