Alkaline Phosphatase Recombinant Rabbit Monoclonal Antibody [SA40-00]
cat.: ET1601-21
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: SA40-00
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 57 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Alkaline Phosphatase aa 18-50.
Positive control: Saos-2 cell lysate, HeLa cell lysate, A549 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, HepG2 cell lysate, MG-63 cell lysate, Hela, HepG2, SW480, human liver tissue, human liver carcinoma tissue, human kidney tissue, mouse kidney tissue, mouse bone tissue, rat spleen tissue lysate, rat lung tissue lysate, mouse jawbone tissue.
Subcellular location: Cell membrane, Mitochondrion membrane, Mitochondrion intermembrane space, Extracellular vesicle membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000
1:50
1:50-1:200
1:50
Use at an assay dependent concentration.
Uniprot #: SwissProt: P05186 Human | P09242 Mouse | P08289 Rat
Alternative names: Alkaline phosphatase Alkaline phosphatase placental Alkaline phosphatase placental type Alkaline phosphatase Regan isozyme ALP Alp1 ALPP FLJ61142 Germ-cell alkaline phosphatase nagao isozyme OTTHUMP00000164354 PALP Placental alkaline phosphatase 1 placental heat-stable alkaline phosphatase placental type PLAP PLAP-1 PLAP1 PPB1_HUMAN
Images
ET1601-21_1.jpg Fig1: Western blot analysis of Alkaline Phosphatase on different lysates with Rabbit anti-Alkaline Phosphatase antibody (ET1601-21) at 1/5,000 dilution.

Lane 1: Saos-2 cell lysate (15 µg/Lane)
Lane 2: HeLa cell lysate (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Lane 4: Mouse liver tissue lysate (20 µg/Lane)
Lane 5: Rat liver tissue lysate (20 µg/Lane)

Predicted band size: 57 kDa
Observed band size: 75 kDa

Exposure time: 24 seconds;

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 (ET1601-21) at 1/5,000 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.
ET1601-21_2.jpg Fig2: ICC staining of Alkaline Phosphatase in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-21, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1601-21_3.jpg Fig3: ICC staining of Alkaline Phosphatase in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-21, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1601-21_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Alkaline Phosphatase antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-21, 1/200) for 30 minutes 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.
ET1601-21_5.jpg Fig5: Flow cytometric analysis of Alkaline Phosphatase was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-21, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
ET1601-21_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse jawbone tissue with Rabbit anti-Alkaline Phosphatase antibody (ET1601-21) at 1/200 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 (ET1601-21) at 1/200 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.
ET1601-21_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Alkaline Phosphatase antibody (ET1601-21) at 1/8,000 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 (ET1601-21) at 1/8,000 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.
ET1601-21_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Alkaline Phosphatase antibody (ET1601-21) at 1/8,000 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 (ET1601-21) at 1/8,000 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.