Phospho-c-Fos (S32) Recombinant Rabbit Monoclonal Antibody [PSH21-99]
cat.: HA751815
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell
Clonality: Monoclonal
Clone number: PSH21-99
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 41 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser32 of Human c-Fos.
Positive control: HeLa starved for 16 hours then treated with 200nM TPA for 4 hours cell lysate, HeLa cells starved for 16 hours then treated with 200nM TPA for 4 hours.
Subcellular location: Nucleus, Endoplasmic reticulum, Cytoplasm, cytosol.
Recommended Dilutions:
  WB
  IF-Cell

1:5,000
1:1,000
Uniprot #: SwissProt: P01100 Human
Alternative names: Activator protein 1 AP 1 C FOS Cellular oncogene c fos Cellular oncogene fos FBJ murine osteosarcoma viral (v fos) oncogene homolog (oncogene FOS) FBJ murine osteosarcoma viral oncogene homolog FBJ murine osteosarcoma viral v fos oncogene homolog FBJ Osteosarcoma Virus FOS FOS protein FOS_HUMAN G0 G1 switch regulatory protein 7 G0/G1 switch regulatory protein 7 G0S7 Oncogene FOS p55 proto oncogene c Fos Proto oncogene protein c fos Proto-oncogene c-Fos v fos FBJ murine osteosarcoma viral oncogene homolog
Images
HA751815_1.jpg Fig1: Western blot analysis of Phospho-c-Fos (S32) on different lysates with Rabbit anti-Phospho-c-Fos (S32) antibody (HA751815) at 1/5,000 dilution.

Lane 1: HeLa starved for 16 hours cell lysate
Lane 2: HeLa starved for 16 hours then treated with 200nM TPA for 4 hours cell lysate
Lane 3: HeLa starved for 16 hours then treated with 200nM TPA for 4 hours cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 15 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 55 kDa

Exposure time: 42 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751815) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751815_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells untreated / starved for 16 hours then treated with 200nM TPA for 4 hours labeling Phospho-c-Fos (S32) with Rabbit anti-Phospho-c-Fos (S32) antibody (HA751815) at 1/1,000 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-c-Fos (S32) antibody (HA751815) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.