Raji全细胞裂解物(ab30124)

Q&A (4)

概述

  • 产品名称
    Raji全细胞裂解物
    参阅全部 Raji 细胞裂解液
  • 常规说明


    Protein is isolated from whole tissue homogenates using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70ºC.

  • 经测试应用
    适用于: WBmore details

性能

  • 形式
    Liquid
  • 存放说明
    Store at -80°C. Allow to warm to room temp and agitate gently before aliquotting.
  • 存储溶液
    pH: 7.90
    Constituents: 1% Tergitol-NP40, Potassium chloride, 0.1% Bromophenol blue, 1.2% Beta mercaptoethanol, 2% SDS, HEPES, Sucrose, 10% Glycerol
  • Concentration information loading...
  • 裂解物说明
    Protein is isolated from whole tissue homogenates using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70ºC.
  • 研究领域
  • 背景
    Established in 1963 from the left maxilla of a 12-year-old African boy with Burkitt's lymphoma, it is the first continuous human hematopoietic cell line. It has become an indispensable tool in haematological research and leukaemia and lymphoma studies.

应用

Our Abpromise guarantee covers the use of ab30124 in the following tested applications.

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

应用 Ab评论 说明
WB Use at an assay dependent dilution. Samples need to boiled for 5 minutes and put on ice before use.

文献

ab30124 has not yet been referenced specifically in any publications.

发表研究结果有使用 ab30124？请让我们知道，以便我们可以引用本数据表中的参考文章。

客户评价及客户问答

Q&A
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1-4 of 4 Q&A

Question

Would the following product be expected to have functional proteases
(assuming there are not denaturants in the extraction steps):
Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate - tumor
cell line (ab30124)

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Answer

Thank you for contacting us.


This protein lysate could contain proteases. That’s why we put protease inhibitor cocktail in the protein lyses buffer.


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

vielen Dank für Ihre schnelle Antwort. Wir werden den von Ihnen empfohlenen Isotyp bestellen.
Im Rahmen der Antikörperbestellung möchten wir auch eine Positivkontrolle für den Westernblot (in Form eines Lysats) mitbestellen.
Bei dem IL-9 AK (ab111915) ist auf dem Datasheet angegeben, dass „human fetal liver lysate“ für den Westernblot verwendet wurde. Wie viel µg wurden hier aufgetragen?
Für den IL-9R AK (ab106287) würden wir voraussichtlich „MOLT4 whole cell lysate“ im Westernblot verwenden. Bei einem ausschließlich für Westernblot getesteten IL-9 AK (ab66106) haben Sie dies im Westernblot eingesetzt. Wie viel µg wurden hier auftragen?
Vielen Dank im Voraus.
Mit freundlichen Grüßen

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Answer

Vielen Dank für Ihre Anfrage.

Für ab111915 wurden 20 µg Lysat aufgetragen, für ab66106 waren es10 µg (siehe Datenblatt). Normalerweise sind - je nach Expressionslevel in der Positivkontrolle - 20 bis maximal 50 µg mehr als ausreichend, bei rekombinantem Protein sogar noch weniger.

Wie Sie vielleicht schon gesehen haben, haben wir die entsprechenden Lysate im Katalog:

ab29890: Liver (Human) Tissue Lysate - fetal normal tissue

https://www.abcam.com/index.html?datasheet=29890 (or use the following: https://www.abcam.com/index.html?datasheet=29890).

ab7912: MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate

https://www.abcam.com/index.html?datasheet=7912 (or use the following: https://www.abcam.com/index.html?datasheet=7912).

ab30124: Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate - tumor cell line

https://www.abcam.com/index.html?datasheet=30124 (or use the following: https://www.abcam.com/index.html?datasheet=30124).

Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Question

Dear technical support team: The customer raised an enquiry about ab30124, ab30125, ab30126 this customer wants to know which one is more suitable and could be a positive control of ab52610 (Anti-PLCG 2 (phospho Y1217) antibody [EP1404Y])? According to the online datasheet, the wb result shows “Lane 2: Raji cell lysate with pervanadate”, therefore, she wants to know how to prepare this sample? Could you please offer any suggestion to her? Thanks for your kindly help Best regards

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Answer

Thank you for your enquiry regarding ab52610.

The phosphorylation of PLCG 2 at Tyrosine 1217 needs to be induced to be able to detect this specific modification. As the WB image on the datasheet indicates Raji cell lysate without stimulation will not provide signal for phospho Y1217. It is important to treat the cells with inducer such as pervanadate.

If you need any further assistance in the future, please do not hesitate to contact me.

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Question

We are trying to develop a teaching lab in which students would purify monoclonal antibodies from a cell line, and then verify/quantify their purification by ELISA.   The hybridoma that we have is L243, (produces anti-human MHC class 2), so we are now looking for a source for purified MHC class 2 in order to complete the ELISA.   I found a paper which describes a protocol for purifying MHC class 2 from Raji cells, however we currently do not have this cell line so I was thinking that perhaps if we could find Raji whole cell lysate we may  be able to use it instead.   In the paper I found, a total of 10^7 Raji cells were digested with papain to release surface proteins.  Protease inhibitor is then added and the cells are pelleted leaving the MHC class 2 in the supernatant.  This is followed by dialysis and then the MHC2 protein is conjugated to NHS-activated Sepharose affinity resin.   My questions regarding using the Raji cell lysate are:   -In obtaining the lysate are surface/membrane proteins released and therefore within the lysate itself…..i.e. is there any possibility the surface/membrane proteins are lost from the lysate, (could they still be stuck in the cell membrane which could potentially get spun out and removed from the lysate)?   -How many cells are you starting with in order to get the 100ug of lysate?  The paper that I am trying to adapt is digesting 10^7 Raji cells.   Thank you for any help you can offer.

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Answer

Thank you for contacting Abcam Our protein lysate is called total protein lysate, which means most of the proteins, including membrane and surface proteins, are within the lysate. We don’t really have a cell number to generate 100ug lysate, we can only estimate in this way, assuming 125 million cells as 5 g, and usually there is about 10% lysate, so 500 mg /125million cells, then 100 ug lysate is estimated from 25,000 cells. I have also attached the Abcam protocols book that contains a section on choosing the correct lysis buffer, when trying to capture different parts of the cell. If there is anything else I can help you with, please let me know.  

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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