PA5-79300

antibody from Invitrogen Antibodies

Targeting: GAS6 AXLLG, AXSF, DKFZp666G247, FLJ34709

Immunohistochemistry

Antibody data

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Validation data

Product number: PA5-79300 - Provider product page
Provider: Invitrogen Antibodies
Product name: GAS6 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.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 500 µg/mL
Storage: -20°C

GAS6 protein structure - PA5-79300 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GAS6 in Lane 1: rat brain tissue lysate, Lane 2: mouse brain 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 GAS6 polyclonal antibody (Product # PA5-79300) 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 analysis of GAS 6 in, Lane 1: rat brain tissue lysates, Lane 2: mouse brain 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 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 GAS6 Polyclonal Antibody (Product # PA5-79300) 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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for GAS 6 at approximately 75KD. The expected band size for GAS 6 is at 79KD.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-GAS6 Polyclonal Antibody (Product # PA5-79300) and a 74-79 kDa band corresponding to Growth arrest-specific protein 6 was observed across all the cell lines tested. An additional band was also observed at ~50 kDa in PANC-1 and MCF7 cell lines, which might correspond to a cleaved C-terminal fragment. Whole-cell extracts (30 µg lysate) of PANC-1 (Lane 1), PANC-1 treated with PTI (1X for 4h) (Lane 2), Hep G2 (Lane 3), Hep G2 treated with PTI (1X for 4h) (Lane 4), MIA PaCa-2 (Lane 5), U-87 MG (Lane 6), MCF7 (Lane 7) and BJ (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, (1:4000 dilution)) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of GAS 6 using anti-GAS 6 antibody (Product # PA5-79300) . GAS 6 was detected in a section of A431 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-GAS 6 antibody (Product # PA5-79300) 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Growth arrest-specific protein 6 was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with GAS6 Polyclonal Antibody (Product # PA5-79300) at 1:100 dilution in 0.1% BSA, incubated at 4-degree Celsius overnight, and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.