PA5-79591

antibody from Invitrogen Antibodies

Targeting: LCN2 24p3, NGAL

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-79591

Provider

Invitrogen Antibodies

Product name

NGAL Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL. Positive Control - WB: Mouse Lung Tissue, Mouse Intestine Tissue. IHC: Mouse Spleen tissue, Rat Spleen tissue IHC-F: mouse lung tissue, mouse spleen tissue, rat lung tissue, rat spleen tissue.

Reactivity

Human, Mouse, Rat

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 NGAL in Lane 1: mouse lung tissue lysate, Lane 2: mouse Intestine tissue lysate using 50 µg (reducing conditions) per well. Electrophoresis was performed on 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours and protein was transferred to a nitrocellulose membrane at 150mA for 50-90 minutes. Sample was blocked with 5% Non-fat Milk/TBS for 1.5 hours at room temperature, incubated with NGAL polyclonal antibody (Product # PA5-79591) at a dilution of 0.5 µg/mL (overnight at 4°C), followed by goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10,000. Signal development was performed using a chemiluminescence (ECL) kit.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NGAL Polyclonal Antibody(Product # PA5-79591) and a 22kDa band corresponding to NGAL was observed across all tested cell lines and tissues, and was upregulated in RAW 264.7 upon Thapsigargin treatment. Whole cell extracts (30 µg lysate) of RAW 264.7 (Lane 1), RAW 264.7 (Thapsigargin (500nM, 48 hours))(Lane 2), Rat Liver (Lane 3), Rat Kidney (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 (0.2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of rat lung tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of rat spleen tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of mouse lung tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of mouse spleen tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of rat spleen tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL on paraffin-embedded mouse spleen tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL on paraffin-embedded rat spleen tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NGAL in frozen sections of mouse lung tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NGAL polyclonal antibody (Product# PA5-79591) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of NGAL in A431 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with NGAL Polyclonal Antibody (Product # PA5-79591) 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.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of NGAL in A431 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with NGAL Polyclonal Antibody (Product # PA5-79591) 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.