概述

  • 产品名称
  • 描述
    小鼠多克隆抗体to RED1
  • 宿主
    Mouse
  • 经测试应用
    适用于: WB, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Full length protein corresponding to amino acids 1-741 of Human RED1 (NP_056648.1).

  • 阳性对照
    • Human kidney tissue lysate, A431 cell lysate, RED1 transfected 293T cell line lysate and HeLa cells.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液
    pH: 7.20
    Constituent: 2.68% PBS
  • Concentration information loading...
  • 纯度
    Protein A purified
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab89436 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 a concentration of 1 µg/ml. Predicted molecular weight: 81 kDa.
ICC/IF Use a concentration of 10 µg/ml.

靶标

  • 功能
    Catalyzes the hydrolytic deamination of adenosine to inosine in double-stranded RNA (dsRNA) referred to as A-to-I RNA editing. This may affect gene expression and function in a number of ways that include mRNA translation by changing codons and hence the amino acid sequence of proteins; pre-mRNA splicing by altering splice site recognition sequences; RNA stability by changing sequences involved in nuclease recognition; genetic stability in the case of RNA virus genomes by changing sequences during viral RNA replication; and RNA structure-dependent activities such as microRNA production or targeting or protein-RNA interactions. Can edit both viral and cellular RNAs and can edit RNAs at multiple sites (hyper-editing) or at specific sites (site-specific editing). Its cellular RNA substrates include: bladder cancer-associated protein (BLCAP), neurotransmitter receptors for glutamate (GRIA2 and GRIK2) and serotonin (HTR2C), GABA receptor (GABRA3) and potassium voltage-gated channel (KCNA1). Site-specific RNA editing of transcripts encoding these proteins results in amino acid substitutions which consequently alter their functional activities. Edits GRIA2 at both the Q/R and R/G sites efficiently but converts the adenosine in hotspot1 much less efficiently. Can exert a proviral effect towards human immunodeficiency virus type 1 (HIV-1) and enhances its replication via both an editing-dependent and editing-independent mechanism. The former involves editing of adenosines in the 5'UTR while the latter occurs via suppression of EIF2AK2/PKR activation and function. Can inhibit cell proliferation and migration and can stimulate exocytosis.
  • 组织特异性
    Highly expressed in brain and heart and at lower levels in placenta. Fair expression in lung, liver and kidney. Detected in brain, heart, kidney, lung and liver (at protein level). Isoform 5 is high expressed in hippocampus and colon. Isoform 5 is expressed in pediatric astrocytomas and the protein has a decreased RNA-editing activity. The decrease in RNA editing correlates with the grade of malignancy of the tumors, with the high grade tumors showing lower editing is seen.
  • 序列相似性
    Contains 1 A to I editase domain.
    Contains 2 DRBM (double-stranded RNA-binding) domains.
  • 细胞定位
    Nucleus. Nucleus > nucleolus. Shuttles between nucleoli and the nucleoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ADARB 1 antibody
    • ADARB1 antibody
    • 1700057H01Rik antibody
    • ADAR2 antibody
    • ADAR2a antibody
    • ADAR2a L1 antibody
    • ADAR2a L2 antibody
    • ADAR2a L3 antibody
    • ADAR2b antibody
    • ADAR2c antibody
    • ADAR2d antibody
    • ADAR2g antibody
    • Adarb1 antibody
    • Adenosine deaminase, RNA specific, 2 antibody
    • Adenosine deaminase, RNA specific, B1 (homolog of rat RED1) antibody
    • Adenosine deaminase, RNA specific, B1 (RED1 homolog rat) antibody
    • Adenosine deaminase, RNA specific, B1 antibody
    • AW124433 antibody
    • AW558573 antibody
    • BB220382 antibody
    • D10Bwg0447e antibody
    • Double stranded RNA specific editase 1 antibody
    • Double-stranded RNA-specific editase 1 antibody
    • DRABA2 antibody
    • DRADA2 antibody
    • dsRNA adenosine deaminase antibody
    • EC 3.5.-.- antibody
    • Human dsRNA adenosine deaminase DRADA2 antibody
    • Human dsRNA adenosine deaminase DRADA2b, EC 3.5 antibody
    • OTTHUMP00000115341 antibody
    • OTTHUMP00000115342 antibody
    • RED 1 antibody
    • RED1_HUMAN antibody
    • RNA editase antibody
    • RNA editase 1 antibody
    • RNA editing deaminase 1 antibody
    • RNA editing enzyme 1 antibody
    • RNA editing enzyme 1, rat, homolog of antibody
    • RNA specific adenosine deaminase B1 antibody
    • RNA-editing deaminase 1 antibody
    • RNA-editing enzyme 1 antibody
    see all

图片

  • Anti-RED1 antibody (ab89436) at 1 µg/ml + Human kidney lysate at 50 µg

    Developed using the ECL technique.

    Predicted band size: 81 kDa
    Observed band size: 81 kDa

  • Anti-RED1 antibody (ab89436) at 1 µg/ml + A431 cell lysate at 50 µg

    Developed using the ECL technique.

    Predicted band size: 81 kDa
    Observed band size: 85 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-RED1 antibody (ab89436) at 1 µg/ml

    Lane 1 : RED1 transfected 293T cell lysate
    Lane 2 : Non-transfected 293T cell lysate

    Lysates/proteins at 25 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 81 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?
    Additional bands at: 80 kDa. We are unsure as to the identity of these extra bands.

  • ab89436 at 10 µg/ml staining RED1 in HeLa cells by Immunofluorescence.

文献

ab89436 has not yet been referenced specifically in any publications.

客户评价及客户问答

1-2 of 2 Q&A

Answer

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange the alternative free of charge replacement. I can confirm that 1 vial of ab5408 has been added to your order number [BO] AB-101017. This has Abcam order reference 1072242.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

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Question

Dear technical support team:

This customer has purchase ab89436 (Anti-RED1 antibody) and has conducted the wb with human sample. The results show the expression was doesn’t decrease of knowdown sample; therefore, she wants to ask for your help to modify her experiment step, could you please offer any suggestion to help her?

I also attached her image in this letter and her experiment step as follow:

1. Order details:

Batch number: gr37579-2

Abcamproduct code: ab89436

Antibody storage conditions (temperature/reconstitution etc) 20℃


2. Please describe the problem (high background, wrong band size, more bands, no band etc).

The expression was doesn't decrease of knowdown sample

3. On what material are you testing the antibody in WB?

· Species: human

· What’s cell line or tissue: HeLa cell

· Cell extract or Nuclear extract: Cell extract

· Purified protein or Recombinant protein:


3. The lysate

How much protein was loaded:50 ug

What lysis buffer was used: WCE


What protease inhibitors were used: PI (Roche)

What loading buffer was used:

Phosphatase inhibitors :NaF, Na4VO3, souium pyrophosphate, beta-glycerolphosphate

Did you heat the samples: temperature and time:100℃, 10min

4. Electrophoresis/Gel conditions/ Transfer conditions


Reducing or non reducing gel:Reducing gel

Reducing agent: DTT

Gel percentage :8%

Transfer conditions: (Type of membrane, Protein transfer verified): PVDF,


5. Blocking conditions

Buffer: 1XTBST

Blocking agent: milk, BSA,serum, what percentage: 5% non-fat milk

Incubation time: 1 hr

Incubation temperature: RT


6. Primary Antibody

Species: mouse

Reacts against: human

At what dilution(s) have you tested this antibody: 1/1000
What dilution buffer was used: 5% non-fat milk in 1X TBST
Incubation time: O/N
Incubation temperature: 4℃
What washing steps were done: 5 times, 1X TBST

7. Secondary Antibody

Species: goat-anti-mouse IgG

Reacts against:

At what dilution(s) have you tested this antibody: 1/5000

Incubation time: 1 hr

Wash steps: 5 times, 1X TBST

Fluorochrome or enzyme conjugate: HRP

Do you know whether the problems you are experiencing come from the secondary? no

8. Detection method
ECl, ECl+, other detection method: ECL

9. Did you apply positive and negative controls along with the samples? Please specify. Other antibody and siRNA-treated cells

10. Optimization attempts

How many times have you tried the Western?2 times

Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):

Do you obtain the same results every time e.g. are background bands always in the same place? Yes

What steps have you altered? no

Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly sent will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial. I have been in contact with the originator and theyhave kindly offered toretest this antibody. I can provide you with further information as soon as this has been done.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

In addition, I can suggest you may like to consider the following suggestions as a check for future experiments:

1. For siRNA knockouts, we would suggest it is beneficial to always consider including an endogenous negative control to help assess how well the knock out and antibody procedure are working.

2. We have the following document regarding siRNA experiments on our website which I hope may be helpful to you:
https://www.abcam.com/index.html?pageconfig=resource&rid=12137

Thank you for your cooperation and patience. I look forward to hearing from you with details of how you would like to proceed.

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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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