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    products/reagents/10x-citrate-buffer-ph-60-ab64214.pdf

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10x Citrate Buffer pH 6.0 (ab64214)

  • Datasheet
Reviews (1)Q&A (5)References (8)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)

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Overview

  • Product name

    10x Citrate Buffer pH 6.0
    See all Citrate buffer reagents
  • Tested applications

    Suitable for: IHC-Pmore details
  • General notes

    Citrate buffer pH 6.0 for heat-induced antigen retrieval (HIER) during IHC.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Shipped at Room Temperature. Store at Room Temperature.
  • Storage buffer

    pH: 6.00
    Constituent: Citrate Buffer

    Buffer x10 concentrated.
  • Concentration information loading...
  • Research areas

    • Kits/ Lysates/ Other
    • Tools and Reagents
    • IHC Tools/ Reagents
    • Kits/ Lysates/ Other
    • Kits
    • IHC tools
    • Antigen retrieval reagent
    • Other
  • Relevance

    Citrate buffer is used on formalin-fixed, paraffin-embedded tissue sections mounted on glass slides for target retrieval prior to immunohistochemistry (IHC) procedures. Target retrieval prior to IHC procedures obtains positive results, or results in an increase in staining intensity with many primary antibodies.

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab64214 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
IHC-P
Use at an assay dependent dilution.
Notes
IHC-P
Use at an assay dependent dilution.

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)Jujo T et al. PLoS One (2014) Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemical analysis of mouse lung. Samples were fixed in 10% buffered formalin, paraffinized and sliced at 1.5 µm thick. Antigen retrieval was performed using ab64214 for the deparaffinized slices. Sections were blocked with 2% normal goat serum, PBS(-) and 0.1% Tween20. They were then incubated with the primary antibodies for 1 hour at 4°C and with secondary antibodies for 30 minutes at room temperature. The avidin-biotin-peroxidase complex method with peroxidase streptavidin and the DAB substrate kit ab64238 was performed.

    (A) A resected lung from a mouse sacrificed 28 days after SCL injection.

    (B) Haemotoxylin and Eosin staining showing the tumor composed of a central area with necrosis and a peripheral zone filled with SCLs.

    (C) Immunohistochemical staining for MMP-14 showing positive expression of MMP-14 in the peripheral zone of the tumor and negative in central zone.

    (D) Immunohistochemical staining weak staining of MMP-2 in the peripheral zone. 

     

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 10x Citrate Buffer pH 6.0 (ab64214)Jujo T et al. PLoS One (2014)

    Immunohistochemical analysis of liver tissue section labeling PCNA with ab18197 at 1/250. Antigen retrieval was performed with antigen retrieval buffer, ab64236. A and B, PCNA staining in right and left grafts 1 day after reperfusion. E and F, PCNA staining in right and left grafts 5 day after reperfusion.

    This image was produced using ab64236 (a higher concentration of ab64214 ). Therefore, we expect ab64214 to work as well when diluted to the required concentration. 

     

Protocols

  • 10x Citrate BufferpH 6.0

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (8)

Publishing research using ab64214? Please let us know so that we can cite the reference in this datasheet.

ab64214 has been referenced in 8 publications.

  • Atsumi S  et al. Focal adhesion ribonucleoprotein complex proteins are major humoral cancer antigens and targets in autoimmune diseases. Commun Biol 3:588 (2020). PubMed: 33067514
  • He Y  et al. Tumor immunohistochemistry and preoperative magnetic resonance imaging features predict local recurrence of giant cell tumor of bone following intralesional curettage. Oncol Lett 17:1425-1434 (2019). PubMed: 30675196
  • Ali F  et al. Parvalbumin-Positive Neuron Loss and Amyloid-ß Deposits in the Frontal Cortex of Alzheimer's Disease-Related Mice. J Alzheimers Dis 72:1323-1339 (2019). PubMed: 31743995
  • Dzidziguri D  et al. Determination of The Properties of Rat Brain Thermostable Protein Complex which Inhibit Cell Proliferation. Cell J 19:552-558 (2018). PubMed: 29105389
  • Celebi AR  et al. Evaluation of the 'Hedgehog' signaling pathways in squamous and basal cell carcinomas of the eyelids and conjunctiva. Oncol Lett 12:467-472 (2016). PubMed: 27347166
  • Jujo T  et al. The role of matrix metalloproteinase in the intimal sarcoma-like cells derived from endarterectomized tissues from a chronic thromboembolic pulmonary hypertension patient. PLoS One 9:e87489 (2014). PubMed: 24489925
  • Dai S  et al. Quantification of metallothionein on the liver and kidney of rats by subchronic lead and cadmium in combination. Environ Toxicol Pharmacol 36:1207-16 (2013). PubMed: 24184865
  • Zhang Y  et al. Establishment of animal model of dual liver transplantation in rat. PLoS One 7:e40818 (2012). PubMed: 22829887

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Good product

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
very easy to use and works well
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 14 2020

Question

The tissue I used was rabbit brain tissue. We are specifically looking at cortex. The tissue was harvested via trans cardinal perfusion with PBS and followed by 4% PFA. We then do a 24 hour post fix in the 4% at 4C. After the post fix we put the tissue in HBHS buffer.

Our IHC protocol includes a 30min incubation in NaBH4 to expose the antigens, a 30min incubation in Triton X-100 to lyse the membrane, a 30min incubation in hydrogen peroxide to block endogenous peroxidases and then a 30 min incubation in Image-iT FX to block background staining.

The sections are 200 um thick and we incubate the tissues with the antibody over night at room temp on a shaker.

We do not do an antigen retriever right now but that might be worth looking into.

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Abcam community

Verified customer

Asked on Nov 29 2012

Answer

Thank you for your reply.

Your protocol seems comprehensive, although I would strongly recommend including the antigen retrieval step since this has worked well for other customers. We would recommend citrate buffer pH 6, which we do sell as ab64214.

https://www.abcam.com/10x-citrate-buffer-ph-6-0-ab64214.html

If you're looking for an antigen retrieval protocol, we have some online as well.

https://www.abcam.com/index.html?pageconfig=resource&rid=11488

If you do try the antigen retrieval step and still see no staining, I would be happy to replace or refund the product for you.

I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Nov 29 2012

Question

Hope you are doing well !

Please see below an e-mail, we need to do complete Rabies IHC test... i.e all materials from Antibody diluent to retrival to everything..


Please advice me ASAP.. Also the customer needs specific protocol, as she is very new to IHC.

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Abcam community

Verified customer

Asked on Jul 31 2012

Answer

Thank you for contacting us.

Here is the list with the requested items. However, please be aware that this is not a technical inquiry as all item could be found by using the search application on our website under the "reagents" category. Please let us know if you need help on navigating our website so you can help your customers faster than relying on us.


Regarding the Rabies antibody, I am afraid we do not have what your customer requests: All we have are mouse monoclonal antibodies against this virus, but none produced in goat. Please let me know if this expectable for her, so I can suggest the right primary antibody with the matching biotinylated secondary antibody.


https://www.abcam.com/10x-Citrate-Buffer-pH-60-ab64214.html (or use the following: https://www.abcam.com/10x-Citrate-Buffer-pH-60-ab64214.html).

https://www.abcam.com/10x-EDTA-Buffer-pH-80-ab64216.html (or use the following: https://www.abcam.com/10x-EDTA-Buffer-pH-80-ab64216.html).

https://www.abcam.com/DAB-Substrate-Kit-ab64238.html (or use the following: https://www.abcam.com/DAB-Substrate-Kit-ab64238.html).

https://www.abcam.com/Streptavidin-HRP-ready-to-use-ab64269.html (or use the following: https://www.abcam.com/Streptavidin-HRP-ready-to-use-ab64269.html).

https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html (or use the following: https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html).

https://www.abcam.com/Protein-Block-ab128988.html (or use the following: https://www.abcam.com/Protein-Block-ab128988.html).

https://www.abcam.com/Mayers-Hematoxylin-ab128990.html (or use the following: https://www.abcam.com/Mayers-Hematoxylin-ab128990.html).



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

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Abcam Scientific Support

Answered on Jul 31 2012

Question

I need to develop double IHC analysis on formalin-fixed paraffin-embedded (FFPE) tissue sections of human tumors specimens, and after we have to do laser capture microdissection of positive cells.   I already have two primary antibodies directed against CD44 (produced in mouse) and against ALDH1 (produced in rabbit).   I'm looking for double staining kit (or all of the individual components which analysis needs) with alcohol insoluble chromogens because after IHC and before laser cature microdissection I have to dehydrate the slides.   Thank you for your time and consideration.

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Abcam community

Verified customer

Asked on Feb 21 2012

Answer

Thank you for contacting us.

We do not currently offer double staining IHC kits. Your staining conditions are challenging as, of the two common HRP chromogens, only DAB is alcohol insoluable. While of the common AP chromagens, only BCIP/NBT is insoluable. This means that to do double labeling you will have to do a sequential stain first using one detection system and then with the other.

I have collected a list, with links, of the reagents which you would need for this type of staining.

1) After de-parrafinizing and re-hydrating the samples, perform an antigen retrieval step treating with Cirate buffer pH6.0,https://www.abcam.com/10x-Citrate-Buffer-pH-6-0-ab64214.html, in the microwave or pressure cooker. After performing the antigen retrieval rinse in cold tap water for 10 minutes.

2) Rinse slides in TBST,https://www.abcam.com/20x-TBS-T-with-Tween-20-ab64250.html.

3) Performing a blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

4) Incubate sections with your first primary antibody, for clarity I will assume the CD44 antibody, raised in mouse.

5) Wash with TBST.

6) Block endogenous peroxidases using aHydrogen Peroxide Blocking Reagent,https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html.

7) Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Mouse IgG,https://www.abcam.com/Biotinylated-Goat-anti-Mouse-IgG-H-L-Ready-to-Use-ab64255.html.

8) Wash with TBST.

9) Detect your first antibody usingStreptavidin Peroxidase (Ready to Use)https://www.abcam.com/Streptavidin-Peroxidase-Ready-to-Use-ab64269.html. Note that this does not containa serum or BSA solution as diluent. Serum may contain biotin therefore competing Streptavidin binding with biotinylated secondary antibody, therefore reducing binding activity.

10) Apply your chromogen, DABsubtrate,https://www.abcam.com/DAB-Substrate-Kit-ab64238.html.

11) Wash with TBST.

12)Performing a second blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

13)Wash with TBST.

14)Incubate sections with the your second primary antibody, this time ALDH1, raised in rabbit.

15)Wash with TBST.

16)Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Rabbit IgG,https://www.abcam.com/Biotinylated-Goat-Anti-Rabbit-IgG-H-L-Ready-to-use-ab64256.html

17)Wash with TBST.

18)Detect your first antibody usingStreptavidin Alkaline Phosphatase (Ready to Use)https://www.abcam.com/Streptavidin-Alkaline-Phosphatase-Ready-to-Use-ab64268.html.

19) Apply your second chromogenAlkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use,https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIP-NBT-Ready-to-Use-ab7468.html.

18) Wash with TBST, counterstain if desired.


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

Read More

Abcam Scientific Support

Answered on Feb 21 2012

Question

ab64214 What is the concentration of the citrate in the buffer?

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Abcam community

Verified customer

Asked on Oct 13 2011

Answer

Thank you for your recent telephone enquiry. I can confirm that the concentration of citrate in the buffer is 30 mM. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us. Good luck with your research!

Read More

Abcam Scientific Support

Answered on Oct 13 2011

Question

How would I make a 0.1M solution of this? I am horrible with dilutions and I'm having a hard time figuring this one out...Any help would be greatly appreciated! I'm sorry, I think I actually need 10mM. This is for antigen retrieval step in IHC-P (formalin fixed).

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Abcam community

Verified customer

Asked on Sep 03 2008

Answer

Thank you for your enquiry. Unfortunately, the concentration of 10 X citrate buffer is proprietary and can not be released. However, we can suggest that you make a ten times dilution of the buffer for the antigen retrieval. Simple take 1ml of this solution and dilute with 9ml of deionized water (DW). I hope this information helps. If there is anything else that I can help you with, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Sep 03 2008

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