AFP Recombinant Rabbit Monoclonal Antibody [PSH11-75]
cat.: HA723371
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH11-75
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 69 kDa
Isotype: IgG
Immunogen: Recombinant protein within human AFP aa 1-609.
Positive control: HepG2 cell lysate, Mouse placenta tissue lysate, human placenta tissue, human poorly differentiated hepatocellular carcinoma tissue, mouse embryo liver tissue, rat embryo liver tissue, HepG2.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P

1:2,000
1:100
1:20,000
Uniprot #: SwissProt: P02771 Human | P02772 Mouse | P02773 Rat
Alternative names: Afp AFPD Alpha fetoglobulin Alpha fetoprotein Alpha fetoprotein precursor Alpha-1-fetoprotein Alpha-fetoglobulin Alpha-fetoprotein alpha-fetoprotein, Hereditary persistence of, included FETA FETA_HUMAN Hereditary persistence of alpha fetoprotein HPAFP
Images
HA723371_1.jpg Fig1: Western blot analysis of AFP on different lysates with Rabbit anti-AFP antibody (HA723371) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: Mouse placenta tissue lysate (40 µg/Lane)

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: Lane 1: 25 seconds; Lane 2: 8 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 (HA723371) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723371_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723371) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723371_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human poorly differentiated hepatocellular carcinoma tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723371) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723371_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human well-differentiated hepatocellular carcinoma tissue (negative) with Rabbit anti-AFP antibody (HA723371) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723371) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723371_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse embryo liver tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723371) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723371_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat embryo liver tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723371) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723371_7.jpg Fig7: Immunocytochemistry analysis of HepG2 cells labeling AFP with Rabbit anti-AFP antibody (HA723371) at 1/100 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-AFP antibody (HA723371) at 1/100 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”.