Overview

  • Product name
    Anti-BAFF antibody [Buffy 2]
    See all BAFF primary antibodies
  • Description
    Rat monoclonal [Buffy 2] to BAFF
  • Host species
    Rat
  • Specificity
    Recognizes membrane-bound and soluble human and mouse (although weaker in mouse) BAFF.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Fr, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant soluble protein, corresponding to amino acids 83 - 285 of Human BAFF. It recognizes the TNF-homology domain.

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab16081 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
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab35768 - Rat monoclonal IgM, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration.

Target

  • Function
    Cytokine that binds to TNFRSF13B/TACI and TNFRSF17/BCMA. TNFSF13/APRIL binds to the same 2 receptors. Together, they form a 2 ligands -2 receptors pathway involved in the stimulation of B-and T-cell function and the regulation of humoral immunity. A third B-cell specific BAFF-receptor (BAFFR/BR3) promotes the survival of mature B-cells and the B-cell response.
  • Tissue specificity
    Abundantly expressed in peripheral blood Leukocytes and is specifically expressed in monocytes and macrophages. Also found in the spleen, lymph node, bone marrow, T-cells and dendritic cells. A lower expression seen in placenta, heart, lung, fetal liver, thymus, and pancreas.
  • Sequence similarities
    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    The soluble form derives from the membrane form by proteolytic processing.
    N-glycosylated.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ApoL related ligand TALL 1 antibody
    • B cell Activating Factor antibody
    • B lymphocyte stimulator antibody
    • B-cell-activating factor antibody
    • BAFF antibody
    • BLyS antibody
    • CD 257 antibody
    • CD257 antibody
    • CD257 antigen antibody
    • Delta BAFF antibody
    • Dendritic cell derived TNF like molecule antibody
    • Dendritic cell-derived TNF-like molecule antibody
    • DTL antibody
    • DTL precursor antibody
    • PRO738 antibody
    • soluble form antibody
    • TALL 1 antibody
    • TALL-1 antibody
    • TALL1 antibody
    • THANK antibody
    • TN13B_HUMAN antibody
    • TNF and APOL related leukocyte expressed ligand 1 antibody
    • TNF homolog that activates apoptosis antibody
    • TNF homolog that activates apoptosis NKFB and JNK. antibody
    • TNF- and APOL-related leukocyte expressed ligand 1 antibody
    • TNFSF13B antibody
    • TNFSF20 antibody
    • TNLG7A antibody
    • Tumor necrosis factor (ligand) superfamily member 13b antibody
    • Tumor necrosis factor ligand 7A antibody
    • Tumor necrosis factor ligand superfamily member 13b antibody
    • Tumor necrosis factor ligand superfamily member 20 antibody
    • Tumor necrosis factor like protein ZTNF4 antibody
    • Tumor necrosis factor superfamily member 13B antibody
    • UNQ401 antibody
    • ZTNF4 antibody
    see all

Images

  • Staining of paraffin section of synovial tissue from a patient with Rheumatoid Arthritis. Arrows indicate vessels (20x) staining positive for BAFF using ab16081.

    Tissues are in paraffin sections. Sections are de-waxed and rehydrated, then cooked. Sections are stained in wet boxes, rehydrated and finally prepared as coverslip slides using Eukitt solution.

  • ICC/IF image of ab16081 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab16081, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 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.

  • Overlay histogram showing THP1 cells stained with ab16081 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16081, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgM (mu chain) (ab98368) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgM (ab35768, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Gerzanich V  et al. Salutary effects of glibenclamide during the chronic phase of murine experimental autoimmune encephalomyelitis. J Neuroinflammation 14:177 (2017). Read more (PubMed: 28865458) »
  • Li H  et al. Tripterygium Wilfordii inhibits tonsillar IgA production by downregulating IgA class switching in IgA nephropathy. Oncotarget 8:109027-109042 (2017). Read more (PubMed: 29312588) »
See all 10 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Answer

We have many products in catalogue. Please find below few of these

https://www.abcam.com/cd19-antibody-epr5906-ab134114.html

https://www.abcam.com/cd19-antibody-epr5906-ab134114.html

https://www.abcam.com/cd19-antibody-2e2b6b10-ab31947.html

https://www.abcam.com/baff-antibody-buffy-2-ab16081.html

https://www.abcam.com/baff-antibody-ab117256.html

The compatible secondary antibodies can be found using the below link

https://www.abcam.com/Search?AppliedFacets.FacetProductType=Secondary+antibodies

We have complete set of IHC kits available. Please check the link;

https://www.abcam.com/index.html?pageconfig=resource&rid=12906

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Answer

According to our records, ab16081 and ab16232 was proving difficult to use when staining spleen and aorta tissues presumably in immunohistochemistry and we were in contact in order to help resolve the issue.
Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution. I have attached a questionnaire which will help you to explain the problems encountered with these antibodies and will allow us to investigate the case more closely. Alternatively, you could submit your complaint by filling out the enquiry form found at the following web address:
https://www.abcam.com/index.html?pageconfig=inquiry
If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.
We wish you the best of luck with your research and look forward to a reply.

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Answer

Le laboratoire m'a confirmé que de récents tests ont montré que l'anticorps fonctionne en effet en cytométrie en flux pour le marquage extracellulaire et intracellulaire également.

La fiche technique de l'anticorps [Buffy 2] sera très prochainement mise à jour avec ces nouvelles données.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Answer

Merci de nous avoir contactés.

Il a été décrit que le clone [Buffy 2] ne distingue pas les 2 formes de la BAFF et peut donc reconnaître à la fois la forme soluble et la forme fixée à la membrane.

J'espère que cette information vous sera utile. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Answer

I am sorry to hear that you have experienced difficulties with two of our products in the past. However, because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.
We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.
Regarding the problems encountered with ab16081 and ab16232, if you would like to contact scientific support with information about the protocol used and what difficulties you encountered, we would be happy to help.
If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.
Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.
To find out more about our Abreview system, please see the following link:
https://www.abcam.com/abreviews
I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Question
Answer

Thank you for your reply.

I am sorry that the vials you received did not contain the full amount of the product and I apologize for the inconvenience. As requested, I have issued a free of charge replacement vials of ab16081 and ab16232 with the order number ######.

Please do not hesitate to contact us if you need anything further.

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Answer

Thank you for contacting us.

I have some further information to share with you in regards to the antibodies ab119904, ab79056 and ab86574.

Anti-CD134 / OX40 (ab119904)

This antibody was used in staining of paraffin embedded formalin fixed tissues using sodium citrate buffer (10 mM, pH6) for 15 minutes for antigen retrieval.

Anti-IL17 (ab79056)

This antibody has successfully been used to stain dog lymph node tissue of formalin fixed, paraffin embedded tissue sections again using citrate buffer as the antigen retrieval agent.

Anti-TNFRSF18 (ab86574)

I am seeking further information on what antigen retrieval was used with this antibody and will let you know as soon as I have further details.

As we have discussed previously, it may be that these conditions are not optimal with the tissues you are hoping to use and some optimisation of the protocol will most likely need to be performedin order to achieve the best results.

I have noticed that the antibodies you have chosen have quite different species listed as compatible with the antibody. If you would be able to share with me exactly what samples you intend to use with your staining I can have a look to see if these are the most appropriate antibodies to use or if we have alternatives which may be preferable.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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Question
Answer

Thank you for contacting us.

Your credit note ID is 20547.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for your reply.

Regarding the three options mentioned, product ab65360 is not currently in stock, and it is yet to be confirmed the estimated time of arrival.

Product ab16081 is a good seller with low rate of complaints, but unfortunately there isn’t any WB mage available on its datasheet. Ab16087 has been specifically validated for WB, and contains an image on its datasheet, which can be helpful to users.

In any case, any of the options chosen should work in WB with Human species, as it is specified on the datasheets. In case they didn’t, our Abpromise guarantee would ensure a replacement or credit note in compensation if the complaint is reported within 6 months of purchase.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for your patience with this reply. I finally heard back from the lab as they were looking into this further. The production manager said that IgMs really are pentameric and nothing else, but it is possible that there could potentially be things other than IgMs present in the mixture. There is some difficulty in purifying IgMs, so these antibodies are prepared by precipitating all the proteins from the ascites, and might contain heavy chain portions of the antibody. So while the answer is “The antibody in question is only present as a pentamer,” there might be things that look like (at least parts of) monomers present. So if you run this product in PAGE, you will see a major band corresponding to IgM pentamers, and possibly some additional Fc portions and other proteins that have slipped through the purification process. I hope this information helps. Please contact us with any other questions.  

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1-10 of 13 Abreviews or Q&A

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