710851

antibody from Invitrogen Antibodies

Targeting: CDKN2D INK4D, p19

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

710851

Provider

Invitrogen Antibodies

Product name

p19 INK4d Recombinant Polyclonal Antibody (14HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

14HCLC

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 HEK- 293 (Lane1), HEK- 293 treated with Trichostatin A, (Lane 2), HeLa (Lane 3), HeLa treated with Trichostatin A (Lane 4), HCT 116 (Lane 5), HCT 116 treated with Trichostatin A (Lane 6) (2uM/24hrs). The blots were probed with Anti-P19INK4D Recombinant Rabbit Polyclonal Antibody (Product # 710851, 0.5-1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A clear 18 kDa band corresponding to P19INK4D was observed across cell lines tested. 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).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

P19INK4D was immunoprecipitated using 2 µg of the P19INK4D Recombinant Rabbit Polyclonal Antibody (Product # 710851) from lysate of HeLa cells treated with Trichostatin A (2 uM/24 hours) (Lane 3) using the Dynabeads® Protein A Immunoprecipitation Kit (Product # 10006D).Normal Rabbit IgG was used as a negative IP control (Lane 2). 10% input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using P19INK4D Recombinant Rabbit Polyclonal Antibody (Product # 710851) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).