Anti-PC1/3 抗体 (ab3532)

Reviews (1)Q&A (2)References (2)

Key features and details

  • Rabbit polyclonal to PC1/3
  • Suitable for: IP, WB
  • Reacts with: Mouse
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-PC1/3 antibody
    PC1/3 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to PC1/3

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IP, WBmore details

  • 種交差性

    交差種: Mouse
    交差が予測される動物種: Rat, Cow, Human

  • 免疫原

    Synthetic peptide corresponding to Mouse PC1/3 aa 111-127.
    Sequence:

    SVQKDSALDLFNDPMWN


    (Peptide available as ab5011)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3532 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IP Use at an assay dependent concentration.
WB Use a concentration of 0.4 µg/ml. Detects a band of approximately 87 kDa (predicted molecular weight: 80 kDa).Can be blocked with PC1/3 peptide (ab5011).

ターゲット情報

  • 機能

    Involved in the processing of hormone and other protein precursors at sites comprised of pairs of basic amino acid residues. Substrates include POMC, renin, enkephalin, dynorphin, somatostatin and insulin.

  • 関連疾患

    Defects in PCSK1 are the cause of proprotein convertase 1 deficiency (PC1 deficiency) [MIM:600955]. PC1 deficiency is characterized by obesity, hypogonadism, hypoadrenalism, reactive hypoglycemia as well as marked small-intestinal absorptive dysfunction It is due to impaired processing of prohormones.

  • 配列類似性

    Belongs to the peptidase S8 family. Furin subfamily.

  • 細胞内局在

    Cytoplasmic vesicle > secretory vesicle. Localized in the secretion granules.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • BDP antibody
    • NEC 1 antibody
    • NEC1 antibody
    • NEC1_HUMAN antibody
    • Neuroendocrine convertase 1 antibody
    • PC1 antibody
    • PC3 antibody
    • PCSK1 antibody
    • Prohormone convertase 1 antibody
    • Prohormone convertase 1/3 antibody
    • Prohormone convertase 3 antibody
    • Proprotein convertase 1 antibody
    • SPC3 antibody
    see all

画像

  • Western blot - Anti-PC1/3 antibody (ab3532)
    Western blot - Anti-PC1/3 antibody (ab3532)
    Western blot detection of PC1/3 from mouse pituitary extract using ab3532.
  • Western blot - Anti-PC1/3 antibody (ab3532)
    Western blot - Anti-PC1/3 antibody (ab3532)
    Anti-PC1/3 antibody (ab3532) at 1 µg/ml + Spinal Cord (Mouse) Tissue Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 80 kDa
    Observed band size: 87 + 68 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 35 kDa, 60 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    PC1/3 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The band observed at 68 kDa could potentially be a cleaved form of PC1/3 due to the presence of a 27 amino acid signal peptide and an 83 amino acid propeptide.

参考文献 (2)

ab3532 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab3532 は 2 報の論文で使用されています。

  • Irwin N  et al. Chemical cholecystokinin receptor activation protects against obesity-diabetes in high fat fed mice and has sustainable beneficial effects in genetic ob/ob mice. Biochem Pharmacol : (2012). PubMed: 23085436
  • Gagnon J  et al. Expression of PCSK1 (PC1/3), PCSK2 (PC2) and PCSK3 (furin) in mouse small intestine. Regul Pept 152:54-60 (2009). PubMed: 18706454

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Immunohistochemistry (Frozen sections) abreview for Anti-PC1/3 antibody

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse brain (hypothalamus))
Permeabilization
No
Specification
mouse brain (hypothalamus)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
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Ali Seifinejad

Verified customer

投稿 May 02 2017

Question

A replacement vial for ab3532 would be great. I do believe it was the delay in shipping that is the source of the problem.

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Answer

I have received the following information from the scientist who tested ab3532: "The alignment of the C-terminal peptides shows non cross reactivity to mouse is not surprising since there are not 5 contiguous amino acids that are identical in both sequences of mouse and human". I have updated this information on the datasheet and believe therefore that a replacement vial will not give you better results than the original vial. Would you like to have a replacement vial of another antibody? I look forward to hearing from you to arrange a new free order,

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Question

BATCH NUMBER 90293 ORDER NUMBER 147797 DESCRIPTION OF THE PROBLEM no bands SAMPLE STC-1 cells (mouse intestinal endocrine) lysates PRIMARY ANTIBODY abcam rabbit anti-PC1(ab12275) or abcam rabbit anti-PC2(ab3532), 1:2500 or 1:500, 0.01%tween-20/PBS, 1hr RT or overnight 4C plus 1hr RT washing: 2x 1min, 1x 15min, 3x 5 min DETECTION METHOD ECL Plus (Amersham, cat RPN2132) POSITIVE AND NEGATIVE CONTROLS USED loaded STC-1 lysates ANTIBODY STORAGE CONDITIONS antibody dispensed in 2uL aliquots and stored at -20C SAMPLE PREPARATION lysate buffer: complete mini protease inhibitor (Roche, cat #11836153001) AMOUNT OF PROTEIN LOADED tried 10 and 20ug of total protein lysates per lane ELECTROPHORESIS/GEL CONDITIONS 95C at 5 min, DTT, NuPAGE 4-12% Bis-Tris gel (Invitrogen), 150V 1.5 hrs TRANSFER AND BLOCKING CONDITIONS 10% methanol/NuPAGE transfer buffer, 30 V 1.5 hrs, blocking: 5% skim milk/PBS/ 0.01% tween-20 SECONDARY ANTIBODY goat anti-rabbit HRP conjugated (another company), 1:10,000, diluted in 0.01% tween-20/PBS washing: 2x 1min, 1x 15min, 3x 5 min HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? 1) increased primary antibody conc from 1:2500 to 1:500 2) increased incubation time from 1hr RT to overnight at 4c plus 1hr RT 3) increased secondary antibody conc from 1:10,000 to 1:5,000 ADDITIONAL NOTES To test whether any protein was transferred, I probed the same membrane that did not yield any bands with the anti-PC1/PC2 antibodies, with anti-B tubulin and then HRP conjugated secondary. Using this antibody I was able to detect the appropriate bands for this ubiquitous protein.

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Answer

Thank you very much for your protocol details and your order number. I firstly looked at your protocol and think it is very good. I found references for the presence of both PC1 and PC2 in STC-1 cells (Yoon et al, Endocrinol. 1997 for example) and therefore the problem is certainly not due to low amounts of the protein in your samples. However, we have not determined whether ab12275 does cross react with mouse PC2 and I am currently enquiring to find out the exact homology of the immunogen with the mouse protein; I performed a homology search with the last 15 amino acids which contain the immunogen and found 86% homology. I think therefore that it would be important to determine the exact homology so that we can determine if the problem with ab12275 is a species cross reactivity problem. I secondly looked at the shipping records of your order and indeed found a delay; the package was held by regulatory agency on May 24th and I am worried that the package was stored in a hot area. I would like to offer you a replacement vial of ab3532 already and to wait for the cross reactivity information regarding ab12275, as if it is unlikely to cross react with mouse I would recommend to use a different antibody to detect PC2. I appreciate your patience and will e-mail you as soon as I find out the missing information regarding ab12275,

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