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

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Images

Immunoprecipitation - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533), 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
Multiclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PIPFlow Cyt (Intra)ICC/IF
Human
Tested
Tested
Tested
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500 - 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
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
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

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

Key facts

Isotype
IgG
Form
Liquid
Clonality
Multiclonal
Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
RM1283
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

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

  • - The sensitivity of polyclonal antibodies by recognising multiple epitopes
  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

View our range of recombinant multiclonal antibodies.

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

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.

7 product images

  • Immunoprecipitation - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Immunoprecipitation - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

    Alkaline Phosphatase was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial cell) whole cell lysate with ab322533 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322533 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Saos-2 (human osteosarcoma epithelial cell) whole cell lysate

    Lane 2: ab322533 IP in Saos-2 (human osteosarcoma epithelial cell) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab322533 in Saos-2 whole cell lysate

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

    All lanes: Immunoprecipitation - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533) at 1/30 dilution

    All lanes: Saos-2 (human osteosarcoma epithelial cell) whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 6s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

    Immunohistochemical analysis of paraffin-embedded A Wild-type HAP1 (chronic myelogenous leukemia cell) cell pellet B ALPL knockout HAP1 cell pellet tissue labeling Alkaline Phosphatase with ab322533 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on (A) wild-type HAP1 cell pellet, no staining on (B) ALPL knockout HAP1 cell pellet. The section was incubated with ab322533 for 30mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

    Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling Alkaline Phosphatase with ab322533 at 1/2000 (0.259 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human cervical carcinoma. The section was incubated with ab322533 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

    Immunohistochemical analysis of paraffin-embedded Human adrenal gland tissue labeling Alkaline Phosphatase with ab322533 at 1/2000 (0.259 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human adrenal gland. The section was incubated with ab322533 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Alkaline Phosphatase with ab322533 at 1/2000 (0.259 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on endothelial cells in human spleen. The section was incubated with ab322533 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

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

    Low expression: A549 (PMID: 33858415).

    In Western blot, ab322533 was shown to bind specifically to ALPL. Target of interest was observed at 80 kDa in wild-type HAP1 cell lysates (lane 4) with no signal observed at this size in ALPL knockout cell line (lane 5).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    Exposure time: Lanes 1-3: 6 seconds; Lanes 4-5: 180 seconds

    All lanes: Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533) at 1/1000 dilution

    Lane 1: Saos-2 (human osteosarcoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: Wide-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

    Lane 5: ALPL knockout HAP1 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

    Performed under reducing conditions.

    Observed band size: 80 kDa, 36 kDa

  • Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533), expandable thumbnail

    Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533)

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

    The identity of the lower MW band at approximately 50 kDa is unknown.

    All lanes: Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (ab322533) at 1/1000 dilution

    All lanes: Human liver tissue 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: 180s

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Product protocols

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