Skip to main content

Rabbit Recombinant Monoclonal PPBT antibody. Suitable for IHC-P, WB and reacts with Human samples.

Be the first to review this product! Submit a review

Images

Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (AB305305), expandable thumbnail
  • Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (AB305305), expandable thumbnail
  • Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (AB305305), expandable thumbnail
  • Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (AB305305), expandable thumbnail
  • Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (AB305305), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow Cyt (Intra)ICC/IFIHC-PWB
Human
Not recommended
Not recommended
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended

Not recommended
Not recommended

Species
Human, Rat, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Rat, Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/2000
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Rat
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Mouse
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Rat, Mouse
Dilution info
-
Notes

-

Target data

Function

Alkaline phosphatase that metabolizes various phosphate compounds and plays a key role in skeletal mineralization and adaptive thermogenesis (PubMed:12162492, PubMed:23688511, PubMed:25982064). Has broad substrate specificity and can hydrolyze a considerable variety of compounds: however, only a few substrates, such as diphosphate (inorganic pyrophosphate; PPi), pyridoxal 5'-phosphate (PLP) and N-phosphocreatine are natural substrates (PubMed:12162492, PubMed:2220817). Plays an essential role in skeletal and dental mineralization via its ability to hydrolyze extracellular diphosphate, a potent mineralization inhibitor, to phosphate: it thereby promotes hydroxyapatite crystal formation and increases inorganic phosphate concentration (PubMed:23688511, PubMed:25982064). Acts in a non-redundant manner with PHOSPHO1 in skeletal mineralization: while PHOSPHO1 mediates the initiation of hydroxyapatite crystallization in the matrix vesicles (MVs), ALPL/TNAP catalyzes the spread of hydroxyapatite crystallization in the extracellular matrix (By similarity). Also promotes dephosphorylation of osteopontin (SSP1), an inhibitor of hydroxyapatite crystallization in its phosphorylated state; it is however unclear whether ALPL/TNAP mediates SSP1 dephosphorylation via a direct or indirect manner (By similarity). Catalyzes dephosphorylation of PLP to pyridoxal (PL), the transportable form of vitamin B6, in order to provide a sufficient amount of PLP in the brain, an essential cofactor for enzymes catalyzing the synthesis of diverse neurotransmitters (PubMed:20049532, PubMed:2220817). Additionally, also able to mediate ATP degradation in a stepwise manner to adenosine, thereby regulating the availability of ligands for purinergic receptors (By similarity). Also capable of dephosphorylating microbial products, such as lipopolysaccharides (LPS) as well as other phosphorylated small-molecules, such as poly-inosine:cytosine (poly I:C) (PubMed:28448526). Acts as a key regulator of adaptive thermogenesis as part of the futile creatine cycle: localizes to the mitochondria of thermogenic fat cells and acts by mediating hydrolysis of N-phosphocreatine to initiate a futile cycle of creatine dephosphorylation and phosphorylation (By similarity). During the futile creatine cycle, creatine and N-phosphocreatine are in a futile cycle, which dissipates the high energy charge of N-phosphocreatine as heat without performing any mechanical or chemical work (By similarity).

Alternative names

AP-TNAP, TNS-ALP, TNSALP, Alkaline phosphatase liver/bone/kidney isozyme, Phosphoamidase, Phosphocreatine phosphatase, ALPL

Recommended products

Rabbit Recombinant Monoclonal PPBT antibody. Suitable for IHC-P, WB and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR27506-72
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Alkaline Phosphatase also known as ALP or ALKPHOS is an enzyme that removes phosphate groups from various molecules like nucleotides and proteins. This enzymatic activity requires alkaline conditions with a pH optimum around 8–10. ALP is anchored to the cell membrane and exists in several isoforms each with a mass around 57-70 kDa. Different tissues express specific isoforms such as bone liver kidney and placenta. Osteoblasts for example show high levels of alkaline phosphatase expression often detected by techniques like ALP staining or alkaline phosphatase staining.

Biological function summary

Alkaline phosphatase plays important roles in dephosphorylation processes. Particularly in bone it is associated with osteoblast activity where it is important for bone mineralization. The enzyme is not known to be part of a large protein complex but its activity aids in the conversion of inorganic pyrophosphate into phosphate which promotes the deposition of calcium phosphate in bone. ALP staining and ALP staining osteoblast can be used in laboratory settings for studying the mineralization and function of osteoblasts.

Pathways

Alkaline phosphatase is key in the regulation of the phosphate group levels within the organism particularly in the bone mineralization process. It participates in the vitamin D metabolic pathway which influences calcium and phosphorus homeostasis. Its function has a dynamic relationship with proteins like NPP1 which regulates levels of inorganic pyrophosphate acting counter to alkaline phosphatase's phosphate-producing activity.

Associated diseases and disorders

Defects in alkaline phosphatase activity are linked to conditions like hypophosphatasia and hyperphosphatasia where mineralization processes are disrupted. In hypophosphatasia decreased ALP activity leads to improper bone formation due to accumulation of inorganic pyrophosphate. This interaction is notable alongside proteins such as tissular nonspecific alkaline phosphatase. On the other hand elevated ALP levels often indicate liver or bone disease pathology where increased enzyme levels reflect altered tissue turnover.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Low expression: A549 (PMID: 33858415 ).

    Exposure time: 

    Lane 1: 6 seconds
    Lanes 2 and 3: 180 seconds

    All lanes: Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305) at 1/1000 dilution

    Lane 1: Saos-2 whole cell lysate at 20 µg

    Lane 2: HeLa whole cell lysate at 20 µg

    Lane 3: A549 whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 80 kDa

    Exposure time: 6s

  • Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    ALPL is a glycoprotein of approximately 80 kDa and detected as a 55 kDa band after treated with Protein Deglycosylation MIX II.

    Exposure time: 37 seconds

    All lanes: Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305) at 1/1000 dilution

    Lane 1: Untreated human adrenal gland tisuue lysate at 20 µg

    Lane 2: Human adrenal gland tissue lysate treated with Protein Deglycosylation Mix II at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Observed band size: 55 kDa, 80 kDa

    Exposure time: 37s

  • Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    The samples were run on a Bis-Tris gel. Performed under reducing conditions.

    False colour image of Western blot: Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.

    In Western blot, ab305305 was shown to bind specifically to Alkaline Phosphatase. A band was observed at 75 kDa in wild-type HAP1 cell lysates whereas no signal observed at this size in Alkaline Phosphatase knockout cell line. To generate this image, wild-type and Alkaline Phosphatase knockout HAP1 cell lysates were analyzed. First, samples were run on a SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: Alkaline Phosphatase knockout HAP1 whole cell lysate at 20 µg

    Lane 3: Human adrenal gland at 20 µg

    Secondary

    Lanes 1 - 3: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Lanes 1 - 3: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 75 kDa

  • Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Immunohistochemical analysis of paraffin-embedded human serous ovarian tissue labeling Alkaline Phosphatase with ab305305 at 1/2000 (0.261 µg/ml) followed by ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Positive staining on human serous ovarian carcinoma (PMID: 30979497).

    The section was incubated with ab305305 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

  • Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Alkaline Phosphatase with ab305305 at 1/2000 (0.261 µg/ml) followed by ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Low expression in human spleen lymphocytes with positive staining on vascular endothelium.

    The section was incubated with ab305305 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

  • Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305), expandable thumbnail

    Immunohistochemistry - Anti-Alkaline Phosphatase antibody [EPR27506-72] (ab305305)

    Immunohistochemical analysis of paraffin-embedded human adrenal gland tissue labeling Alkaline Phosphatase with ab305305 at 1/2000 (0.261 µg/ml) followed by ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Positive staining on human adrenal gland (PMID: 31563571).

    The section was incubated with ab305305 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Leica DS9800 (Bond® Polymer Refine Detection) kit.

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com