Overview

  • Product name

    Anti-ADAMTS2/NPI antibody
  • Description

    Rabbit polyclonal to ADAMTS2/NPI
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Human ADAMTS2/NPI (C terminal). Synthetic peptide corresponding to the C-terminal sequence of Human ADAMTS2/ NPI. The sequence differs from the related rat and mouse sequences by three amino acids.

  • Positive control

    • Rat brain tissue lysate, Rat heart tissue lysate, Rat cardiac muscle tissue. IF/ICC: PANC-1 cell line
  • General notes

    Protein previously labeled as ADAMTS2.

Properties

Applications

Our Abpromise guarantee covers the use of ab125226 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 135 kDa. The detection limit is approximately 1ng/lane under non-reducing and reducing conditions.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function

    Cleaves the propeptides of type I and II collagen prior to fibril assembly. Does not act on type III collagen. May also play a role in development that is independent of its role in collagen biosynthesis.
  • Tissue specificity

    Expressed at high level in skin, bone, tendon and aorta and at low levels in thymus and brain.
  • Involvement in disease

    Defects in ADAMTS2 are the cause of Ehlers-Danlos syndrome type 7C (EDS7C) [MIM:225410]. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS7C is marked by extremely fragile tissues, hyperextensible skin and easy bruising. Facial skin contains numerous folds, as in the cutis laxa syndrome.
  • Sequence similarities

    Contains 1 disintegrin domain.
    Contains 1 peptidase M12B domain.
    Contains 1 PLAC domain.
    Contains 4 TSP type-1 domains.
  • Domain

    The spacer domain and the TSP type-1 domains are important for a tight interaction with the extracellular matrix.
  • Post-translational
    modifications

    The precursor is cleaved by a furin endopeptidase.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links

  • Alternative names

    • A disintegrin and metalloproteinase with thrombospondin motifs 2 antibody
    • A disintegrin like and metalloprotease (reprolysin type) with thrombospondin type 1 motif 2 antibody
    • ADAM metallopeptidase with thrombospondin type 1 motif 2 antibody
    • ADAM TS 2 antibody
    • ADAM TS2 antibody
    • ADAM-TS 2 antibody
    • ADAM-TS2 antibody
    • ADAMTS 3 antibody
    • ADAMTS-2 antibody
    • ADAMTS2 antibody
    • ATS2_HUMAN antibody
    • EC 3.4.24.14 antibody
    • EDS VIIB antibody
    • EDS VIIC antibody
    • hPCPNI antibody
    • NPI antibody
    • PC I NP antibody
    • PC I-NP antibody
    • PCINP antibody
    • PCPNI antibody
    • pNPI antibody
    • Procollagen I N proteinase antibody
    • Procollagen I N-proteinase antibody
    • Procollagen I/II amino propeptide processing enzyme antibody
    • Procollagen I/II amino propeptide-processing enzyme antibody
    • Procollagen N endopeptidase antibody
    • Procollagen N-endopeptidase antibody
    see all

Images

  • All lanes : Anti-ADAMTS2/NPI antibody (ab125226) at 1 µg/ml

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat heart tissue lysate

    Predicted band size: 135 kDa

  • ICC/IF image of ab125226 stained PANC-1 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab125226, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab125226, at 1µg/mL, staining paraffin embedded Rat cardiac muscle tissue by immunohistochemistry (IHC-P).

References

This product has been referenced in:

  • Jana S  et al. Disparate Remodeling of the Extracellular Matrix and Proteoglycans in Failing Pediatric Versus Adult Hearts. J Am Heart Assoc 7:e010427 (2018). Read more (PubMed: 30371322) »
  • Matsuura-Hachiya Y  et al. Recovery of extracellular matrix components by enalapril maleate during the repair process of ultraviolet B-induced wrinkles in mouse skin. Biochem Biophys Rep 4:180-186 (2015). Read more (PubMed: 29124203) »
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jun 06 2016

Application
Western blot
Sample
Mouse Tissue lysate - other (Rat quad, mouse skeletal muscle, HOJ 18ºC)
Gel Running Conditions
Reduced Denaturing (Hand-cast gel (4% stacking section and 12% running section))
Loading amount
45 µg
Specification
Rat quad, mouse skeletal muscle, HOJ 18ºC
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 14 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (bone cells, Saos2)
Permeabilization
Yes - 0.3 % Triton-X-100
Specification
bone cells, Saos2
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Prof. Feray Kockar

Verified customer

Submitted Aug 15 2013

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. It is regrettable the results have not been succesful and I fully understand your disappointment.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB and in human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I notice you have been in contact with us before back in July regarding this matter and were in corresopndence with my colleague Anja. the following reply was sent to you. I hope you had recieved this? I woudl appreciate if you could confirm if the suggestions provided have been tried? It is also particularly important for us to know the order number and date of purchase, as requested, for our records.

Thank you for your cooperation. I look forward to hearing from you with the further requested details.

PREVIOUS REPLY:
Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.
I would like to reassure you that ab125226 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab125226.
1.) I can recommend to use a positive control that is not a cell line. Due to extensive culturing cell line often exhibit artifacts. As positive control I can suggest brain or heart tissue lysates form eat or human. Maybe a second cell line?
2.) Since all bands are smaller than expected I suggest to check the protease inhibitors that are used.
3.) To achieve the most specific signal and the best saturation of the antibody I suggest to incubate the primary antibody over night at 4C.
I would also appreciate if you can confirm some further details:
What is the lot umber of the vial you are using? When was it ordered?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab125226 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab125226.

1.) I can recommend to use a positive control that is not a cell line. Due to extensive culturing cell line often exhibit artifacts. As positive control I can suggest brain or heart tissue lysates form eat or human. Maybe a second cell line?

2.) Since all bands are smaller than expected I suggest to check the protease inhibitors that are used.

3.) To achieve the most specific signal and the best saturation of the antibody I suggest to incubate the primary antibody over night at 4C.

I would also appreciate if you can confirm some further details:

What is the lot umber of the vial you are using? When was it ordered?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

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