1. Antibodies
  2. Homo sapiens
  3. FOXP2

ENCAB000AGQ

Antibody against Homo sapiens FOXP2

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Abcam
Product ID
ab16046
Lot ID
AB-2B
Characterized targets
FOXP2 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Antigen description
Raised against C-terminal 127 amino acids of FOXP2 of human origin

Characterizations

FOXP2 (Homo sapiens)
Method: immunoblot
Attachment PDF Icon
not reviewed
Caption
Western blot protocol: Protein extracts from whole cell lysates of PFSK-1 were loaded on a 7.5% acrylamide gel and separated with a Bio-Rad PROTEAN II xi system. After separation, the samples were transferred to a nitrocellulose membrane. Blotting was performed with primary antibody and secondary HRP-conjugated antibody. Visualization was achieved using ECL Plus detection kit (GE Healthcare). Results: We found that the two antibodies raised against antigen 1, AB-1A and AB-1B, recognized FOXP2 on Western blots of PFSK-1 cell lysates, but were not specific to FOXP2. In contrast, Western blots probed with antibodies AB-2A and AB-2B (raised against antigen 2, the C-terminal 127 amino acids of FOXP2) produced only one band at the expected size of 80 kDa (Figure 1). Figure 1 - FOXP2 immunoblot: western blot with, left to right, FOXP2 antibodies AB-1A, AB-1B, AB-2A, and AB-2B in lysates of PFSK-1. FOXP2 band is indicated at ~80 kDa. AB-2B antibody (fourth lane) is considered positive and subsequently used for ChIP-seq (see validation 2 below).
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
FOXP2 (Homo sapiens)
Method: knockdown or knockout
Attachment PDF Icon
not reviewed
Caption
siRNA protocol: Briefly, shRNAs against FOXP2 were designed using RNAi OligoRetriever. Oligos were purchased as complete hairpin sequences and cloned into the lentiviral vector pSicoR-PGK-puro. Cell culture supernatants from transfected 293T cells were used to infect PFSK-1 cells, followed by selection with puromycin. Cells were harvested for western blotting five days post-infection. Figure Legend: FOXP2 knockdown in PFSK-1 cells assayed by western blot using FOXP2 antibody AB-2B. Upper: Lanes 1-6 (left to right): shRNA1; shRNA2; shRNA3; negative control shRNA; empty lentiviral vector; no lentiviral vector. Lower: β-actin control.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB