711748

antibody from Invitrogen Antibodies

Targeting: IDH2

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711748

Provider

Invitrogen Antibodies

Product name

IDH2 Recombinant Polyclonal Antibody (8 HCLC)

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

8 HCLC

Vial size

100 µg

Concentration

0.5 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 analysis was performed on Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1) HEK-293 (Lane 2), MCF-7 (Lane 3), U-87 MG (Lane 4), LNCaP (Lane 5) and tissue extracts of Rat Kidney (Lane 6), Mouse Skeletal Muscle (Lane 7) and Mouse Heart (Lane 8). The blots were probed with Anti-IDH2 Recombinant Rabbit Polyclonal Antibody (Product # 711748, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). 45, 51 kDa band corresponding to IDH2 was observed respectively across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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 IDH2 was achieved by transfecting Hep G2 cells with IDH2 specific validated siRNA (Silencer® select Product # s225). Western blot analysis (Fig a) was performed using whole cell extracts from IDH2 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-IDH2 Recombinant Rabbit Polyclonal Antibody (Product # 711748, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 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 IDH2.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1) HEK-293 (Lane 2), MCF-7 (Lane 3), U-87 MG (Lane 4), LNCaP (Lane 5) and tissue extracts of Rat Kidney (Lane 6), Mouse Skeletal Muscle (Lane 7) and Mouse Heart (Lane 8). The blots were probed with Anti-IDH2 Recombinant Rabbit Polyclonal Antibody (Product # 711748, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). 45, 51 kDa band corresponding to IDH2 was observed respectively across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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).