JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB322533

Anti-Alkaline Phosphatase antibody [RM1283]

  • 20ul selling size
  • BOND RX™ Validated
  • Recombinant
  • RabMAb
  • KO Validated
  • What is this?

Be the first to review this product! Submit a review

|

(0 Publication)

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

View Alternative Names

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

7 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533)
  • IHC-P

Supplier Data

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)
  • IHC-P

Supplier Data

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)
  • IHC-P

Supplier Data

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)
  • IHC-P

Supplier Data

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

Immunoprecipitation - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533)
  • IP

Supplier Data

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)(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 (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) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533)
  • WB

Supplier Data

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 (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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa,36 kDa

false

Western blot - Anti-Alkaline Phosphatase antibody [RM1283] (AB322533)
  • WB

Supplier Data

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1283

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

WB, IP, IHC-P

applications

Immunogen

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

style
left-column

Complementary Products

Assay Kits

AB83369

Alkaline Phosphatase Assay Kit (Colorimetric)

Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369)

  • 0
  • 0
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/500 - 1/2000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Mouse": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }
style
left-column

Product details

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.

style
left-column

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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
style
left-column
style
left-column
style
left-column

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
style
left-column

Target data

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).
See full target information Alkaline phosphatase, tissue-nonspecific isozyme
style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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