Table 2 Dilutions of primary and secondary antibodies used for immunoblotting detection of proteins involved in hypoxia and p53 activation in human and mouse cancer cell lysates.

From: MDM2 inhibitors, nutlin-3a and navtemadelin, retain efficacy in human and mouse cancer cells cultured in hypoxia

Protein target

Primary antibody

Secondary antibody

1° antibody (clone or catalog)

Host species

Company

Dilution of 1º

2° antibody

Host species

Company

Dilution of 2º

HIF-1α (Hs)

610959

Mouse

BD biosciences

1: 500

Anti-mouse HRP

Goat

Dako

1: 1000

p53 (Hs)

DO-1

Mouse

In house32

1: 3000

Anti-mouse HRP

Goat

Dako

1: 2000

p21 (Hs)

2947

Rabbit

Cell signaling technology

1: 1000

Anti-rabbit HRP

Swine

Dako

1: 2000

HIF-1α (m)

NB100-479

Rabbit

Novus biologics

1: 2000

Anti-rabbit HRP

Swine

Dako

1: 2000

p53 (m)

ab246223

Rabbit

Abcam

1: 1000

Anti-rabbit HRP

Swine

Dako

1: 2000

p21 (m)

ab188224

Rabbit

Abcam

1: 1000

Anti-rabbit HRP

Swine

Dako

1: 2000

  1. HIF-1α hypoxia inducible factor 1α, Hs human, m mouse.