MA5-29624

antibody from Invitrogen Antibodies

Targeting: TXN TRX

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-29624

Provider

Invitrogen Antibodies

Product name

Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

This product is preservative free. It is recommended to add sodium azide to avoid contamination (final concentration 0.05%-0.1%). Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire. This antibody has specificity for Human Thioredoxin/TXN.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

116

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624) at 1:2,000 dilution. Lane A: Hela Whole Cell Lysate, Lane B: THP-1 Whole Cell Lysate, Lane C: A431 Whole Cell Lysate. Lysates/proteins at 30 μg per lane. Secondary Goat Anti-Rabbit IgG H&L (DyLight™ 800) at 1:10,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 12 kDa. Observed band size: 14 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Thioredoxin 1 was achieved by transfecting HeLa with Thioredoxin 1 specific siRNAs (Silencer® select Product # s1 ,s2). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Thioredoxin 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624, 1:2,000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000). 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 Thioredoxin 1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624) and a ~12 kDa band corresponding to Thioredoxin 1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), Hep G2 (Lane 3), THP-1 (Lane 4), A-431 (Lane 5), HEK-293 (Lane 6), A549 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:2,000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Thioredoxin 1 was performed using 70% confluent log phase HeLa 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 Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624, 1:100) 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, 1:2,000), 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 cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Thioredoxin 1 Immunoprecipitation using: Lane A: 0.5 mg Hela Whole Cell Lysate, Lane B: 0.5 mg 293T Whole Cell Lysate 2 µL with Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624) and 15 µL of 50 % Protein G agarose. Primary antibody: Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116), at 1:100 dilution. Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 13 kDa. Observed band size: 13 kDa.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Thioredoxin 1 Immunoprecipitation using: Lane A: 0.5 mg Hela Whole Cell Lysate, Lane B: 0.5 mg 293T Whole Cell Lysate 2 µL with Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116) (Product # MA5-29624) and 15 µL of 50 % Protein G agarose. Primary antibody: Thioredoxin 1 Recombinant Rabbit Monoclonal Antibody (116), at 1:100 dilution. Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 13 kDa. Observed band size: 13 kDa.