Overview

  • Product name
    Anti-Collagen III antibody [FH-7A]
    See all Collagen III primary antibodies
  • Description
    Mouse monoclonal [FH-7A] to Collagen III
  • Host species
    Mouse
  • Tested applications
    Suitable for: Dot blot, ELISA, IHC-Fr, WB, IHC-P, ICC/IF, Indirect ELISAmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Full length native protein (purified) (Human).

  • Positive control
    • IHC: Human skin sections. WB: recombinant collagen III.
  • General notes

    Type III collagen, [a1(III)]3 ,is an approx. 300 kDa molecule, found predominantly in skin, blood vessels, liver, placenta, tongue, and thymus. Collagen type III forms cofibrils with type I and/or V collagens in a number of tissues of mesenchymal origin, such as skin, tendon, ligaments, and bone. This collagen type is involved, directly or indirectly in several genetic diseases, including Ehlers-Danlos type IV disease.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab6310 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
Dot blot Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-Fr 1/4000.
WB 1/100 - 1/1000.
IHC-P 1/600.
ICC/IF Use at an assay dependent concentration. PubMed: 25136258
Indirect ELISA Use at an assay dependent concentration.

Target

  • Function
    Collagen type III occurs in most soft connective tissues along with type I collagen.
  • Involvement in disease
    Defects in COL3A1 are a cause of Ehlers-Danlos syndrome type 3 (EDS3) [MIM:130020]; also known as benign hypermobility syndrome. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS3 is a form of Ehlers-Danlos syndrome characterized by marked joint hyperextensibility without skeletal deformity.
    Defects in COL3A1 are the cause of Ehlers-Danlos syndrome type 4 (EDS4) [MIM:130050]. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS4 is the most severe form of the disease. It is characterized by the joint and dermal manifestations as in other forms of the syndrome, characteristic facial features (acrogeria) in most patients, and by proneness to spontaneous rupture of bowel and large arteries. The vascular complications may affect all anatomical areas.
    Defects in COL3A1 are a cause of susceptibility to aortic aneurysm abdominal (AAA) [MIM:100070]. AAA is a common multifactorial disorder characterized by permanent dilation of the abdominal aorta, usually due to degenerative changes in the aortic wall. Histologically, AAA is characterized by signs of chronic inflammation, destructive remodeling of the extracellular matrix, and depletion of vascular smooth muscle cells.
  • Sequence similarities
    Belongs to the fibrillar collagen family.
    Contains 1 fibrillar collagen NC1 domain.
    Contains 1 VWFC domain.
  • Post-translational
    modifications
    Proline residues at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.
    O-linked glycan consists of a Glc-Gal disaccharide bound to the oxygen atom of a post-translationally added hydroxyl group.
  • Cellular localization
    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha 1 type III collagen antibody
    • Alpha1 (III) collagen antibody
    • CO3A1_HUMAN antibody
    • COL 3A1 antibody
    • COL3A1 antibody
    • Collagen alpha 1(III) chain antibody
    • Collagen alpha-1(III) chain antibody
    • Collagen III alpha 1 chain precursor antibody
    • Collagen III alpha 1 polypeptide antibody
    • Collagen type III alpha 1 (Ehlers Danlos syndrome type IV autosomal dominant) antibody
    • Collagen type III alpha 1 antibody
    • Collagen type III alpha 1 chain antibody
    • Collagen type III alpha antibody
    • Collagen, fetal antibody
    • EDS4A antibody
    • Ehlers Danlos syndrome type IV, autosomal dominant antibody
    • Fetal collagen antibody
    • Type III collagen antibody
    see all

Images

  • All lanes : Anti-Collagen III antibody [FH-7A] (ab6310) at 1/500 dilution

    Lane 1 : Reduced, denaturated Collagen III protein
    Lane 2 : Native Collagen III protein

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Goat anti-mouse HRP at 1/2000 dilution

    Developed using the ECL technique.

    Exposure time: 1 minute
  • Staining of formalin-fixed, paraffin-embedded rat skin with 1:4,000 ab6310 using biotin/ExtrAvidin®-Peroxidase.

  • ab6310 at 1/600 diltuion staining preovulatory follicle and whole ovary tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Antigens were retrieved by boiling with an antigen unmasking solution for 20 min at 120ºC in an autoclave and then cooled down in water for 5 minutes. The tissue sections were formaldehyde fixed and incubated with the antibody for 1 hour. An alkaline phosphatase conjugated antibody was used as the secondary. The image shows a section of whole preovulatory follicle. Staining for collagen type III is seen in the theca interna cell layer. No staining in the granulosa cells.

    See Abreview

  • Staining of frozen rat skin sections with 1:8,000 ab6310 using biotin/ExtrAvidin®-Peroxidase.

References

This product has been referenced in:
  • Fang CL  et al. Liposome-Encapsulated Baicalein Suppressed Lipogenesis and Extracellular Matrix Formation in Hs68 Human Dermal Fibroblasts. Front Pharmacol 9:155 (2018). Read more (PubMed: 29559910) »
  • Molina-Molina M  et al. Anti-fibrotic effects of pirfenidone and rapamycin in primary IPF fibroblasts and human alveolar epithelial cells. BMC Pulm Med 18:63 (2018). Read more (PubMed: 29703175) »
See all 73 Publications for this product

Customer reviews and Q&As

1-10 of 34 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (femoral vein)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
Yes - Triton X
Specification
femoral vein
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 23 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
skin tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted May 24 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH6
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted May 24 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted May 24 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
ELISA
Sample
Human Purified protein (purified protein and cell culture supernatant)
Specification
purified protein and cell culture supernatant
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Type
Sandwich (Capture)

Patricia Carulla

Verified customer

Submitted Jun 28 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (First trimester human decidua)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: 0.1% (W/V) Trypsin
Permeabilization
No
Specification
First trimester human decidua
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde

Ruhul Choudhury

Verified customer

Submitted Feb 13 2017

Answer

Thank you for contacting us. We offer either replacement or refund. As you have already received compensation in the form of a replacement antibody, you would be ineligible for a refund. Please note that your replacement product is covered by our Abpromise guarantee. We are happy provide scientific support at any time. If you are using the product in species and applications listed on the datasheet and contact us within six months of purchase, we are also happy to replace or refund the product should we not be able to help you to resolve the issue. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

Read More

Question

Thank you for your detailed answer about the anti-collagen III antibody ab6310.

That's a pity indeed that the validity on pig was initially a mistake but these things happen. I am a regular customer who appreciate very much your policy about customer feedback and reviewing, and I'll submit an abreview about IHC-Fr in pig skin after performing the few more tests discussed.

In return, to answer your questions:

- From what I've read and optimized so far on frozen sections with different kind of antibodies, the antigen retrieval step is usually not needed as the fixation time is very short.

I don't have any pronase, and I'm not sure it's worth for me buying some only for a test, with no insurance the antibody will work. I can try fixing in acetone instead or formalin see if it changes something.

- I will follow your advice and block with only donkey serum or BSA but not both at the same time

- The secondary antibody works well, I've written reviews about ab90395 (collagen I) and ab6328 (fibronectin EDA), where I used the same anti-mouse Cy2 secondary antibody on similar pig skin frozen sections with good results.

- I have performed the procedure on a couple of separate samples, and I did it twice on independent batches before contacting the technical service, to ensure the problem was from the antibody and not from my experimental procedure.

I’d like to try the reference ab23445, as I also would like to do some western blotting. If this second one doesn't work, will I still be eligible to try the other one, or for a refund?

Thank you very much

Best regards

Read More
Answer

Thank you foryour understanding and cooperation. The details provided enable us to closely monitor the quality of our products.

I hope the suggestions may proove useful with ab6310. It may be worth trying citrate antigen retreival with the PFA fixationif you have the reagents. It is unusual to use antigen retreival with IHC-Fr, however laboratory tests do indicate that this can sometimes improve the results.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issuedthe alternativefree of charge replacement of ab23445with the order number 1132685.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. The ab23445 is tested and covered by the guarantee for IHC-Fr, WB and pig samples. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question

Hello,

I have purchased the anti-collagen III antibody ab6310 a few months ago (order #1076197).

However, after several attemps to optimise the experimental conditions, I haven't managed to make it work properly in IHC-Fr on pig skin cryosections



Here are the conditions I have used for IHC-Fr:

Pig skin cryosections, 12um thick, embedded in OCT

Fixation in either formaldehyde 4% in PBS, or neutral buffer formalin for 10min at RT

Permeabilisation in PBS 0.1% triton for 10min at RT

Blocking in PBS, 5% donkey serum, 1%BSA, 0.01% tween for 1h at RT

Incubation of primary antibody ab6310, diluted 1/500 in PBS , 1% donkey serum, 0.1%BSA, 0.01% tween overnight at 4C

Secondary antibody: Cy2 donkey anti mouse 1/100 for 1h at RT

Mounting in Prolong mounting media

(this protocol worked fine for the collagen I antibody ab90395 for which I submitted a review)

Results: No positive staining was observed in the dermis, either in unwounded skin or in the wound bed (where there should be even more Collagen III).

Another of your anti-col III antibodies ab7778 displays a strong positive staining in human dermis in IHC-P, that’s the kind of signal I was expected.

Could you tell me the experimental conditions you have been using to validate it in IHC-Fr (I was surprised to read a line about some antigen retrieval with pronase, which is usually not necessary with frozen sections, but maybe that’s needed here. However nothing is precised on the first page, in the applications section. Would you have specific technical advices to help me make it work?

Also, in your scientific support section about this antibody, I have just read that this antibody wasn’t actually experimentally validated on pig samples but only predicted to work, based on sequence similarity. I wish I had seen it before so I could have at least asked for the testing offer while purchasing it.

Have you got any feedback of these customers who were working on pig tissue, did they manage to make it work?

I also saw that you offered them to try other antibodies references that may work on pig, have you got more data on these other antibodies?



I look forward to hearing from you

Best regards

Read More
Answer

Thank you for taking the time to contact us with this information. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I have investigated our records for ab6310 and have contacted our source for this antibody. Regrettably,I can confirm that this antibody has not been tested on pig. I am not sure why this information was originally on our datasheet. The datasheet has been updated with the correct information and I sincerely apologize for any confusion. The quality of our antibodies and datasheets is very important to us, as is any feedback and data we receive. It is regrettable that the information we had on this particular antibody was not of the standard both we and our customers expect from our products.

In this case I would be pleased to provide you with an immediatecredit note, free alternative replacement or refund.

Alternatively, as the immunogen for the antibody has high alignment to the pig protein, you may like to consider trying a few suggestions first that I hope may help improve the results.I would also appreciate if you can confirm some further details, to help with our quality monitoring:

1. As requested, we have some information regarding the antigen retreival: ab6310 is only tested on unfixed frozen sections in-house, and the antigen retrieval method is 0.1% pronase for 10 minutes. However we also have feedback from other researchers using this antibody successfully in IHC on FFPE tissue, and heat-mediated antigen retrieval for 20 minutes in pH 6 citrate buffer should work as well.

2. We recommend not to mix blocking agents in one experiment. Try using 2% serum or 2% BSA alone to block.

3. Is the current vial of secondary antibody working well with other primary antibodies?

4. Try some fresh samples.

I hope this information is helpful. We do have the following collagen III antibodies that are tested in IHC-Fr and pig. You may wish to consider trying one of these as a replacement:

ab24119 Rabbit polyclonal Anti-Collagen III antibody
Tested in: ELISA, Dot, IHC-Fr
Reacts with: Human, Pig
https://www.abcam.com/index.html?datasheet=24119 (or use the following: https://www.abcam.com/index.html?datasheet=24119).

ab23445Mouse monoclonalCollagen III antibody [1E7-D7/Col3]
Tested in: WB, ELISA, IHC-Fr
Reacts with: Human, Rat, Dog, Kangaroo, Pig
https://www.abcam.com/index.html?datasheet=23445 (or use the following: https://www.abcam.com/index.html?datasheet=23445).


Should the suggestions not improve the results, please do not hesitate to contact me again with details of how you would like to proceed.

Read More

Question
Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unserem Wissen nach wurde ab7778 bisher noch nicht inSchwein und Schafund ab6310 nicht in Schaf getestet.

Falls Sie dies selbst testen möchten, kann ich Ihnen zurzeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert des Antikörpers abdecken, und gegen eine erneute Bestellung von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte,welchen Antikörper Sietesten möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in Schaf (und/ oder Schwein).

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einihn mit einer neuen Bestellung verrechnen lassen (halten Sie bei der Bestellung bitte den Rabattcode). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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1-10 of 34 Abreviews or Q&A

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