702047

antibody from Invitrogen Antibodies

Targeting: SERPINA1 A1A, A1AT, AAT, alpha-1-antitrypsin, alpha1AT, PI, PI1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

702047

Provider

Invitrogen Antibodies

Product name

alpha-1 Antitrypsin Recombinant Rabbit Monoclonal Antibody (8H10L18)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Dog, Pig, Cat

Antibody clone number

8H10L18

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

A1AT was immunoprecipitated using 5 µg of the Anti-A1AT Recombinant Rabbit Monoclonal Antibody (Product # 702047) from Condition medim of Hep G2 (Lane 2) using the Dynabeads® Protein A Immunoprecipitation Kit (Product # 10006D). Normal Rabbit IgG was used as a Isotype control (Lane 1). Western blot analysis was performed using Anti-A1AT Recombinant Rabbit Monoclonal Antibody (Product # 702047, 2 µg/mL). A 55 kDa band corresponding to alpha-1 antiTrypsin/A1AT was observed in IP elute. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extract (30 µg lysate) of Hep G2 (Lane 1), Hep G2 conditioned media (Lane 2), HT-29 (Lane 3) and HT-29 conditioned media (Lane 4).The blots were probed with Anti-alpha-1 Antitrypsin Recombinant Rabbit Monoclonal Antibody (Product # 702047, 0.5 - 1 µg/mL). A 55 kDa band corresponding to alpha-1 Antitrypsin was observed in both cell lysates and condition media tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of A1AT was achieved by transfecting HT29 cells with A1AT specific siRNA (Silencer® select Cat # s10458 and s10459). Western blot analysis (Fig a) was performed using Whole cell extract from the A1AT knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-A1AT Recombinant Rabbit Monoclonal Antibody (Product # 702047, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to A1AT.