Product: Alkaline Phosphatase Antibody
Catalog: DF6225
Description: Rabbit polyclonal antibody to Alkaline Phosphatase
Application: WB IHC IF/ICC
Cited expt.: WB, IHC, IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 57kDa; 57kD(Calculated).
Uniprot: P05186
RRID: AB_2838191

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(88%), Bovine(88%), Horse(88%), Sheep(88%), Rabbit(100%), Dog(88%)
Clonality:
Polyclonal
Specificity:
ALPL Antibody detects endogenous levels of total ALPL.
RRID:
AB_2838191
Cite Format: Affinity Biosciences Cat# DF6225, RRID:AB_2838191.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AKP2; Alkaline phosphatase liver/bone/kidney; Alkaline phosphatase liver/bone/kidney isozyme; Alkaline phosphatase tissue nonspecific isozyme; Alkaline phosphatase, tissue-nonspecific isozyme; Alkaline phosphomonoesterase; Alpl; AP TNAP; AP-TNAP; APTNAP; BAP; FLJ40094; FLJ93059; Glycerophosphatase; HOPS; Liver/bone/kidney type alkaline phosphatase; MGC161443; MGC167935; PHOA; PPBT_HUMAN; Tissue non specific alkaline phosphatase; Tissue nonspecific ALP; TNAP; TNSALP;

Immunogens

Immunogen:

A synthesized peptide derived from human ALPL, corresponding to a region within the internal amino acids.

Uniprot:
Gene(ID):
Description:
There are at least four distinct but related alkaline phosphatases: intestinal, placental, placental-like, and liver/bone/kidney (tissue non-specific). The product of this gene is a membrane bound glycosylated enzyme that is not expressed in any particular tissue and is, therefore, referred to as the tissue-nonspecific form of the enzyme. Alkaline phosphatases form various dimers in vivo. This antibody can bind the four mentioned alkaline phosphatases.
Sequence:
MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Rabbit
100
Pig
88
Horse
88
Bovine
88
Sheep
88
Dog
88
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

This isozyme plays a key role in skeletal mineralization by regulating levels of diphosphate (PPi).

PTMs:

N-glycosylated.

Subcellular Location:

Cell membrane>Lipid-anchor.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Family&Domains:

Belongs to the alkaline phosphatase family.

Research Fields

· Metabolism > Metabolism of cofactors and vitamins > Thiamine metabolism.

· Metabolism > Metabolism of cofactors and vitamins > Folate biosynthesis.

· Metabolism > Global and overview maps > Metabolic pathways.

References

1). Supramolecular Hydrogel with Ultra-Rapid Cell-Mediated Network Adaptation for Enhancing Cellular Metabolic Energetics and Tissue Regeneration. Advanced Materials, 2024 (PubMed: 38295393) [IF=27.4]

Application: IF/ICC    Species: Human    Sample: hMSCs

Figure 7. The dynamic HA-ADA hydrogel enhances the osteogenic differentiation. D) Representative images and E) quantified intensity (n = 20) of ALP (green) immunostaining of the hMSCs in the hydrogels.

2). Targeted disruption of PRC1.1 complex enhances bone remodeling. Nature Communications, 2025 [IF=15.7]

3). Osteoblast-Derived ECM1 Promotes Anti-Androgen Resistance in Bone Metastatic Prostate Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2025 (PubMed: 39563492) [IF=15.1]

Application: IHC    Species: human    Sample:

Figure 8 Validation of ECM1/ENO1/MAPK signaling axis in patients with bmCRPC. A) Representative images (left) of PCa PDOs treated with Veh (PBS), ECM1 (200 ng mL−1), or ECM1 (200 ng mL−1) combined with either DMSO or PhAH (1 µM) in the presence of ENZ (10 µM). B) Assessment of PDOs growth by changes in organoid diameter (right; Scale bar, 50 µm). C) IHC staining of ALP, ECM1, ENO1, and p-ERK1/2 in ENZ-untreated (n = 12) or ENZ-treated (n = 22) bmCRPC patients tissue (Scale bars, 250 µm and 50 µm). D-G) Staining index quantification of ALP, ECM1, p-ERK1/2 expression and relative ENO1 membrane/cytoplasm cell number as indicated in C. H) Percentage of specimens showing ENZ-untreated or treated in relation to the expression levels of ALP, ECM1, p-ERK1/2 and the membrane or cytoplasm level of ENO1. I-K) Spearman correlation analysis between the expression levels of ALP, relative ENO1 membrane/cytoplasm cell number, and p-ERK1/2 with ECM1 expression. L) Representative mIHC staining images of ALP (red, osteoblast marker), EPCAM (yellow, PCa marker), and ENO1 (green) in human bmCRPC tissues (Scale bars, 100 µm and 25 µm). M) Quantification of ALP, EPCAM, and relative ENO1 membrane/cytoplasm intensity per cell in L). N) WB analysis of MEK, p-MEK, ERK1/2, and p-ERK1/2 expression in prostate cancer cells extracted from human bmCRPC tissue with or without ECM1 treatment (200 ng mL−1). GAPDH was used as the loading control. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Application: IF/ICC    Species: human    Sample:

Figure 8 Validation of ECM1/ENO1/MAPK signaling axis in patients with bmCRPC. A) Representative images (left) of PCa PDOs treated with Veh (PBS), ECM1 (200 ng mL−1), or ECM1 (200 ng mL−1) combined with either DMSO or PhAH (1 µM) in the presence of ENZ (10 µM). B) Assessment of PDOs growth by changes in organoid diameter (right; Scale bar, 50 µm). C) IHC staining of ALP, ECM1, ENO1, and p-ERK1/2 in ENZ-untreated (n = 12) or ENZ-treated (n = 22) bmCRPC patients tissue (Scale bars, 250 µm and 50 µm). D-G) Staining index quantification of ALP, ECM1, p-ERK1/2 expression and relative ENO1 membrane/cytoplasm cell number as indicated in C. H) Percentage of specimens showing ENZ-untreated or treated in relation to the expression levels of ALP, ECM1, p-ERK1/2 and the membrane or cytoplasm level of ENO1. I-K) Spearman correlation analysis between the expression levels of ALP, relative ENO1 membrane/cytoplasm cell number, and p-ERK1/2 with ECM1 expression. L) Representative mIHC staining images of ALP (red, osteoblast marker), EPCAM (yellow, PCa marker), and ENO1 (green) in human bmCRPC tissues (Scale bars, 100 µm and 25 µm). M) Quantification of ALP, EPCAM, and relative ENO1 membrane/cytoplasm intensity per cell in L). N) WB analysis of MEK, p-MEK, ERK1/2, and p-ERK1/2 expression in prostate cancer cells extracted from human bmCRPC tissue with or without ECM1 treatment (200 ng mL−1). GAPDH was used as the loading control. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

4). Liquid-Liquid Phase Separation-Mediated Cellular-Scale Compartmentalization of Hydrogel Covalent Cross-Linking Promotes Microtubule-Based Mechanosensing. Journal of the American Chemical Society, 2025 (PubMed: 40252026) [IF=14.5]

5). Delivery of therapeutic miRNAs using nanoscale zeolitic imidazolate framework for accelerating vascularized bone regeneration. Chemical Engineering Journal, 2022 [IF=13.3]

6). Polydatin accelerates osteoporotic bone repair by inducing the osteogenesis-angiogenesis coupling of bone marrow mesenchymal stem cells via the PI3K/AKT/GSK-3β/β-catenin pathway. International journal of surgery (London, England), 2025 (PubMed: 39248296) [IF=12.5]

7). Polydatin accelerates osteoporotic bone repair by inducing the osteogenesis-angiogenesis coupling of bone marrow mesenchymal stem cells via the PI3K/AKT/GSK-3β/β-catenin pathway. International journal of surgery (London, England), 2024 (PubMed: 39248296) [IF=12.5]

Application: WB    Species: Rat    Sample:

Figure 1. POL stimulated BMSCs proliferation and osteogenic differentiation. (A) POL chemical structure; (B) BMSCs surface markers were detected by flow cytometry; (C) The result of CCK-8 assay (n = 5); (D-E) The result of CFU assay (n = 5); (F) Images of ALP staining (scale bar = 250 μm); (G) Quantification of ALP staining (n = 5); (H) Images of ARS staining (scale bar = 250 μm); (I) Quantification of ARS staining (n = 5); (J-K) The protein expression levels of OCN, RUNX2, ALP, and COL1A1 were evaluated by western blot (n = 3); (L-M) Immunofluorescence staining of OCN and ALP (scale bar = 200 μm). Data were presented as mean ± SEM. Compared with control group: * P < 0.05, **P < 0.01, ***P < 0.001. Compared with 1 μM group: # P < 0.05, ##P < 0.01, ###P < 0.001.

8). circRNA422 enhanced osteogenic differentiation of bone marrow mesenchymal stem cells during early osseointegration through the SP7/LRP5 axis. Molecular Therapy, 2022 (PubMed: 35642253) [IF=12.1]

Application: WB    Species: Rat    Sample: BMSCs

Figure 3 circRNA422 interfering inhibited osteogenic differentiation of BMSCs (A) ALP staining to evaluate the effect of circRNA422− on the ALP activity of BMSCs on osteogenic days 3 and 7. (B) Protein levels of OCN to evaluate the effect of circRNA422− on mineralization of BMSCs on osteogenic days 14 and 21. (C) Quantitative real-time PCR analysis showed that the osteogenic mRNA expressions levels of SP7, LRP5, ALP, OCN, BSP, and OPN were downregulated by circRNA422− compared with Con313 on osteogenic days 3 and 7 (n = 3, ∗p < 0.05). (D) WB analysis showed that the osteogenic protein expressions levels of ALP, LRP5, and SP7 were reduced by circRNA422− compared with Con313 on osteogenic days 3 and 7 (n = 3, ∗p < 0.05).

9). Biomimetic hydroxyapatite coating on the 3D-printed bioactive porous composite ceramic scaffolds promoted osteogenic differentiation via PI3K/AKT/mTOR signaling pathways and facilitated bone regeneration in vivo. Journal of Materials Science & Technology, 2023 [IF=11.2]

10). Micro or nano: Evaluation of biosafety and biopotency of magnesium metal organic framework-74 with different particle sizes. Nano Research, 2020 [IF=9.5]

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