Native Human Collagen VI protein (ab7538)

Submit a review Q&A (7)References (2)

製品の詳細

特性

Our Abpromise guarantee covers the use of ab7538 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

    Western blot

    Immunoprecipitation

    ELISA

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

  • 製品の状態

    Liquid

  • 備考

    This product is free from other collagens, human serum proteins and non-collagen extracellular matrix proteins. This product reacts with anti-Collagen Type VI. Reaction with anti-Collagen I, II, III, IV or V is negligible (typically less than 1% cross reactivity was detected by ELISA).

  • Concentration information loading...

前処理および保存

  • 保存方法および安定性

    Shipped at 4°C. Store at +4°C. Diluted stock solutions should be used immediately and not be frozen. Please see notes section. Store undiluted.

    Preservative: 0.01% Sodium azide
    Constituent: 3% Acetic acid

関連情報

  • 別名

    • Alpha 1 (VI) chain (61 AA)
    • CO6A1_HUMAN
    • COL6A1
    • COL6A2
    • COL6A3
    • collagen 6
    • Collagen alpha 2(VI) chain
    • Collagen alpha 3(VI) chain
    • Collagen alpha-1(VI) chain
    • collagen six
    • Collagen type VI alpha 1
    • Collagen type VI alpha 2
    • Collagen type VI alpha 3
    • Collagen VI alpha 1 polypeptide
    • Collagen VI alpha 2 polypeptide
    • Collagen VI alpha 3 polypeptide
    • CollagenVI
    • Human mRNA for collagen VI alpha 2 C terminal globular domain
    • OPLL
    • PP3610
    see all

  • 機能

    Collagen VI acts as a cell-binding protein.

  • 関連疾患

    Defects in COL6A1 are a cause of Bethlem myopathy (BM) [MIM:158810]. BM is a rare autosomal dominant proximal myopathy characterized by early childhood onset (complete penetrance by the age of 5) and joint contractures most frequently affecting the elbows and ankles.
    Defects in COL6A1 are a cause of Ullrich congenital muscular dystrophy (UCMD) [MIM:254090]; also known as Ullrich scleroatonic muscular dystrophy. UCMD is an autosomal recessive congenital myopathy characterized by muscle weakness and multiple joint contractures, generally noted at birth or early infancy. The clinical course is more severe than in Bethlem myopathy.

  • 配列類似性

    Belongs to the type VI collagen family.
    Contains 3 VWFA domains.

  • 翻訳後修飾

    Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.

  • 細胞内局在

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

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (2)

ab7538 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。

ab7538 は 2 報の論文で使用されています。

  • Holm Nielsen S  et al. A Fragment of Collagen Type VI alpha-3 chain is Elevated in Serum from Patients with Gastrointestinal Disorders. Sci Rep 10:5910 (2020). PubMed: 32245981
  • Veidal SS  et al. MMP Mediated Degradation of Type VI Collagen Is Highly Associated with Liver Fibrosis - Identification and Validation of a Novel Biochemical Marker Assay. PLoS One 6:e24753 (2011). PubMed: 21935455

レビューと Q&A

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1-7 of 7 Abreviews or Q&A

Question

Can you also tell me, whether HUMAN COLLAGEN TYPE VI PROTEIN (ab7538, Lot No. GR107403) is present as tropocollagen with loose telopeptides or as atelocollagen with no telopeptides (collagen monomers)?

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Answer

I have contacted the laboratories who confirmed that the collagen is not assessed specifically for telopeptides. Lower molecular weight subunit strands are not expected in the unreduced collagen.

When the collagen has been loaded on 4-20% gradient gels, only the reduced collagen sample was observed to show bands, the non-reduced sample was too large to run on the gel.

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Question


Can you please tell me, if the collagen in this soloutions is present as triple helixes, microfibrils, fibrils or fibers? What size are they?

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Answer


I can confirm the collagen is present as triple helices. The size is not determined but the native protein is normally too large to run in standard 4-8% polyacrylamide gels.

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Question


I have purchased the isolate. However, I seem to have
a problem detecting the peptide on a western blot or using elisa.
I have loaded 500 ng of the isolate right after receiving it and ran a
Western blot which did not show a very strong signal for ColVI. An
over-exposed membrane showed two very weak bands at around 130 and 150
kD. Col VI should have bands for alpha 1 and alpha 2 at around 140 kD
and for alpha 3 at around 240 kD. I have used a monoclonal COL6A1/2/3
Antibody from another company which we have been
succesfully using in the past to detect Col VI. The positive control
yielded good results on the blot and I had much stronger signal from
nonspecific binding in control than from the isolate.
In addition, I have also tried coating maxisorp plates with 10 and 20
ug/ml of the isolate for elisa (using the same antibody as for
western) and could not detect any signal. The reagents have been
tested on other samples and work properly.
1. What are your suggestions on why I cannot detect the isolate?
2. Could you please send me the image of the Western and all
supporting data from testing the ColVI isolate, indicating which bands
should represent ColVI?
3. I would also like to ask if you have any antibodies which you
recommend for detecting your isolate. The antibody from abcam which is
recommended on your website, however, does not seem to give a very
strong and specific signal according to the image which is on the
website.
Sincerely,

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Answer

Thank you for keeping me updated about the results, and also for your patience while I've been in touch with the lab that produces this protein.

I do have a couple of suggestions that will hopefuly improve the results, but I was unfortunately not able to obtain any data or images from the production lab.

I also have a couple of additional questions that might be helpful for optimization:

1)Were standard reduced, denatured conditions used in the Western blot protocol?
2) What positive control was used in WB?
3) Was a Coomassie or other gel stain done before transfer to the membrane?
4) Do you have any images that you could send to me?

Here is the ELISA protocol that is used to quantify collagen:

1) Coat the plate with unlabeled anti-collagen (type specific) at 10 ug/ml.
2) Use biotinylated anti-collagen (same specificity) at a concentration of 0.1 ug/ml.
3) Dissolve the collagen firstly in 0.5 M Sodium Acetate.
4) Dilute the collagen in 0.05 M Tris Cl, 1.0 M Sodium Chloride, pH. 7.4.
5) Perform serial dilutions of the collagen using the Tris/NaCl buffer.
6) Perform all steps at 4° C using overnight incubations.
7) All buffers are to be at 4° C.
8) Use Streptavidin-HRP for detection.

I don't have the full protocol that was used in WB, however my contact at the lab recommendsnative, non-denaturing conditions (no heat, reducing agents or SDS) in WB. Bands are most frequently reported between 80 and 120 kDa. The antibody you have was raised against the native protein so the affinity should be higher under these conditions.

With this protein, the anti-collagen VI antibodies ab6588 and ab6583 are recommended for detection, however the collagen VI antibody that you have should work as well. We do fully guarantee this protein to work as specified on the datasheet, so if you would prefer a replacement, credit, or refund, please let me know and I'll be happy to arrange this for you.

I hope that this information will be useful, and I look forward to hearing from you. Please let me know if you have further questions or if there is anything else that we can do for you.

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Question

Dear sir/madame,
Does the collagen VI (ab 7538) isolate contain all A1, A2, and A3
chains? Is the protein in native conformation?
Sincerely,

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Answer

Thank you for contacting us with your question.

The collagen VI protein Ab7538 is a native protein, and it is purified full length protein isolated from human placenta. Thus it will contain the three subunits A1, A2, and A3.

I hope this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

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Question

I recently purchased collagen VI (ab7538) from your company. Do you have non-pepsin treated collagen VI available for purchase?

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Answer

Thank you for contacting Abcam regarding our Collagen VI proteins.


I have confirmed with the laboratory that both proteins on our catalog have been pepsin-treated.


I hope this information is helpful and I apologize for any inconvenience this may cause. Please do not hesitate to contact us if you have any additional questions or concerns.

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Question

Has this protein be treated with pepsin? What band size should I expect on a Western blot?

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Answer

Thanks for your call yesterday and for your patience while I have been in touch with the lab regarding your enquiry.

This protein has been treated by limited pepsin digestion.Bands most often appear between 80-120 kDa and vary depending on sample preparation and gel type.Collagen antibodies often have higher affinity for the non-denatured protein, so it is common to run a non-denaturing gel with this protein.

I hope this information is helpful, but please let me know if you have any further questions or if there is anything else that we can do for you.

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Question

I have performed a Western blot using the collagen VI that I have ordered in your company and I see 3 bands between 75 -50 kDa ( using a polyclonal antibody against col VI). My question is if this collagen is pepsinized? Normaly, I should see the alpha 1 and alpha 2 chains around 140 KDa and alpha 3 chain around 240 kDa. I am looking forward to hearing from you. Best Regards,

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Answer

I'm afraid I was unable to find out if this collagen was pepsinised. However, we do not recommend to use reduced conditions of SDS-PAGE for collagens and I would suggest to run a native (non-denaturing, non-dissociating) PAGE which should give you the appropriate band sizes. I hope this recommendation will help, please do not hesitate to contact me again if you still have problems with native conditions,

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