PI3K p110α antibody | knockout validation | Cell Signaling Technology 4249

This is a knockout-validated antibody summary, based on four publications "Tumor-intrinsic PIK3CA represses tumor immunogenecity in a model of pancreatic cancer" [4] (western blot, Figure s1a in the article), "A PI3K p110β-Rac signalling loop mediates Pten-loss-induced perturbation of haematopoiesis and leukaemogenesis" (Figure 1) [1], "Rac1-mediated membrane raft localization of PI3K/p110β is required for its activation by GPCRs or PTEN loss" (Figure 2) [2], and "The Effector TepP Mediates Recruitment and Activation of Phosphoinositide 3-Kinase on Early Chlamydia trachomatis Vacuoles" (Figure 3) [3]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

PI3K p110α antibody | knockout validation | Cell Signaling Technology 4249 figure 1
Figure 1. From Figure S1 in [1]. Western blot of lysates of spleen from wild-type and PI3K p110α -/- mice probed with anti-p110α antibody. Vinculin serves as a loading control.
Antibody information

Rabbit monoclonal

Company: Cell Signaling Technology

Antibody: PI3K p110α

Catalog number: 4249

Summary: Rabbit monoclonal antibody raised against a peptide corresponding to residues surrounding Asp520 of human p110α.

Supplier recommended for WB, IP and IHC.

Reacts with human, mouse, rat and cow p110α.

Article "A PI3K p110β-Rac signalling loop mediates Pten-loss-induced perturbation of haematopoiesis and leukaemogenesis"
Validation Method

Western blot (Figure 1).

PI3K p110α antibody | knockout validation | Cell Signaling Technology 4249 figure 2
Figure 2. Wt, p110α,β-KO (DKO), DKO+p110α-wt and DKO+p110β-wt MEFs were analyzed in immunoblots for expression of p110α and p110β. Actin was used as loading control. From [2].
Sample

Lysates of spleen from wild-type and PI3K p110α -/- mice.

Blocking agent

TBS containing 5% milk powder.

Primary incubation

Anti-p110α antibody diluted 1:1000.

Secondary incubation

LI-COR 800 conjugated anti-rabbit IgG.

Detection

Odyssey detection system (LI-COR).

PI3K p110α antibody | knockout validation | Cell Signaling Technology 4249 figure 3
Figure 3. HeLa cells, and Hela cells with knockout of PIK3KCA, CRKL, or CRKI-II were examined with respective antibodies. Tubulin was used as positive controls. From [3].
Article "Rac1-mediated membrane raft localization of PI3K/p110β is required for its activation by GPCRs or PTEN loss"
Validation Method

Western blot (Figure 2).

Sample

Wt, p110α,β-KO (DKO), DKO+p110α-wt and DKO+p110β-wt MEFs.

Blocking agent

5% non-fat milk (LabScientific, Highlands, New Jersey) in TBS for 30 min.

Primary incubation

Diluted in TBS containing 5% milk and 0.1% Tween-20 overnight at 4°C.

Secondary incubation

IRDye 800CW goat anti-rabbit secondary antibodies (Li-Cor, Lincoln, Nebraska) diluted in 5% milk-TBS for 1.5 hr at RT.

Detection

Odyssey CLx Scanner.

References
  1. Yuzugullu H, Baitsch L, Von T, Steiner A, Tong H, Ni J, et al. A PI3K p110β-Rac signalling loop mediates Pten-loss-induced perturbation of haematopoiesis and leukaemogenesis. Nat Commun. 2015;6:8501 pubmed publisher
  2. Cizmecioglu O, Ni J, Xie S, Zhao J, Roberts T. Rac1-mediated membrane raft localization of PI3K/p110? is required for its activation by GPCRs or PTEN loss. elife. 2016;5: pubmed publisher
  3. Carpenter V, Chen Y, Dolat L, Valdivia R. The Effector TepP Mediates Recruitment and Activation of Phosphoinositide 3-Kinase on Early Chlamydia trachomatis Vacuoles. mSphere. 2017;2: pubmed publisher
  4. Sivaram N, McLaughlin P, Han H, Petrenko O, Jiang Y, Ballou L, et al. Tumor-intrinsic PIK3CA represses tumor immunogenecity in a model of pancreatic cancer. J Clin Invest. 2019;130: pubmed publisher