Anti-BNIP3L/NIX 抗体 (ab8399)

Reviews (3)Q&A (7)References (22)

Key features and details

  • Rabbit polyclonal to BNIP3L/NIX
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-BNIP3L/NIX antibody
    BNIP3L/NIX 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to BNIP3L/NIX

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, ICC/IF, IHC-Pmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Rat

  • 免疫原

    Synthetic peptide corresponding to Human BNIP3L/NIX aa 77-92.
    Sequence:

    DAQHESGQSSSRGSSH


    Database link: O60238
    (Peptide available as ab8431)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • K562 cell lysatecan be used as positive control and the approximate 40kDa top band represents monomer of Bnip3L/NIX (4).

  • 特記事項

     

    Members in the Bcl-2 family are critical regulators of apoptosis by either inhibiting or promoting cell death. Bcl-2 homology 3 (BH3) domain is a potent death domain. BH3 domain containing pro-apoptotic proteins, including Bad, Bax, Bid, Bik, Hrk, Nip3, and Bim, form a growingsubclass of the Bcl-2 family. A novel BH3 domaincontaining protein was recently identified and designated Bnip3L/NIX, Bnip3a, and Nix (for Nip3-like protein X) (Matsushima, M. et al.; Yasuda, M. et al.; Chen, G, et al.).Bnip3L/Bnip3a/Nix is a homolog of the E1B 19K/Bcl-2 binding and pro-apoptotic protein Bnip3. Over expression of Bnip3L/NIX induces apoptosis (Yasuda, M. et al.; Chen, G, et al.). Bnip3L/NIX interacts withand overcomes suppresses by Bcl-2 and Bcl-xL. Bnip3L/NIX is localized in mitochondria. The messenger RNA of Bnip3L/NIX is ubiquitously expressed in human tissues (Matsushima, M. et al.; Yasuda, M. et al.). Bnip3L/NIX and Bnip3 form a new subfamily of the pro-apoptotic mitochondrial proteins.

     This product was previously labelled as BNIP3L

     

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab8399 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 40 kDa. To reduce possible background load less sample and or use more diluted antibody.
ICC/IF Use at an assay dependent concentration. See Abreview.
IHC-P Use a concentration of 2 µg/ml.

ターゲット情報

  • 機能

    Induces apoptosis. Interacts with viral and cellular anti-apoptosis proteins. Can overcome the suppressors BCL-2 and BCL-XL, although high levels of BCL-XL expression will inhibit apoptosis. May function as a tumor suppressor. Inhibits apoptosis induced by BNIP3.

  • 配列類似性

    Belongs to the NIP3 family.

  • 翻訳後修飾

    Undergoes progressive proteolysis to an 11 kDa C-terminal fragment, which is blocked by the proteasome inhibitor lactacystin.

  • 細胞内局在

    Nucleus envelope. Endoplasmic reticulum. Mitochondrion. Membrane.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Adenovirus E1B19k binding protein B5 antibody
    • Adenovirus E1B19K-binding protein B5 antibody
    • BCL2/adenovirus E1B 19 kd protein interacting protein 3a antibody
    • BCL2/adenovirus E1B 19 kDa protein interacting protein 3A antibody
    • BCL2/adenovirus E1B 19 kDa protein-interacting protein 3-like antibody
    • BCL2/adenovirus E1B 19 kDa protein-interacting protein 3A antibody
    • BCL2/adenovirus E1B 19kDa interacting protein 3 like antibody
    • BNI3L_HUMAN antibody
    • BNIP3a antibody
    • BNIP3H antibody
    • BNIP3L antibody
    • BNIP3L protein antibody
    • NIP3 like protein X antibody
    • NIP3-like protein X antibody
    • NIP3L antibody
    • NIX antibody
    see all

画像

  • Western blot - Anti-BNIP3L/NIX antibody (ab8399)
    Western blot - Anti-BNIP3L/NIX antibody (ab8399)
    All lanes : Anti-BNIP3L/NIX antibody (ab8399) at 1 µg/ml

    Lane 1 : K562 whole cell lysate with absence of immunogenic peptide
    Lane 2 : K562 whole cell lysate with presence of immunogenic peptide


    The approximate 40 kDa top band represents monomer of Bnip3L/NIX.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3L/NIX antibody (ab8399)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3L/NIX antibody (ab8399)

    ab8399 at 2µg/ml staining BNIP3L/NIX in human kidney tissue by IHC 

  • Immunocytochemistry/ Immunofluorescence - Anti-BNIP3L/NIX antibody (ab8399)
    Immunocytochemistry/ Immunofluorescence - Anti-BNIP3L/NIX antibody (ab8399)

    Immunofluorescence of Bnip3L/NIX in Human Kidney cells using ab8399 at 10 ug/ml.

参考文献 (22)

ab8399 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab8399 は 22 報の論文で使用されています。

  • Zhu N  et al. Berberine Protects Against Simulated Ischemia/Reperfusion Injury-Induced H9C2 Cardiomyocytes Apoptosis In Vitro and Myocardial Ischemia/Reperfusion-Induced Apoptosis In Vivo by Regulating the Mitophagy-Mediated HIF-1a/BNIP3 Pathway. Front Pharmacol 11:367 (2020). PubMed: 32292345
  • Czernik M  et al. Deregulated Expression of Mitochondrial Proteins Mfn2 and Bcnl3L in Placentae from Sheep Somatic Cell Nuclear Transfer (SCNT) Conceptuses. PLoS One 12:e0169579 (2017). PubMed: 28076382
  • Qi N  et al. Multiple truncated isoforms of MAVS prevent its spontaneous aggregation in antiviral innate immune signalling. Nat Commun 8:15676 (2017). PubMed: 28607490
  • Toschi P  et al. Evidence of Placental Autophagy during Early Pregnancy after Transfer of In Vitro Produced (IVP) Sheep Embryos. PLoS One 11:e0157594 (2016). WB . PubMed: 27326761
  • Wu HM  et al. Hypoxia-induced autophagy mediates cisplatin resistance in lung cancer cells. Sci Rep 5:12291 (2015). WB ; Human . PubMed: 26201611
View all Publications for this product

レビューと Q&A

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1-10 of 10 Abreviews or Q&A

Immunocytochemistry abreview for Anti-BNIP3L/NIX antibody

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Mar 13 2018

Immunocytochemistry abreview for Anti-BNIP3L/NIX antibody

 Average
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Rat Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Feb 28 2018

Question

Hello,

As for the quantity of protein loaded per lane, I used several different loadings ranging from 2ug/uL to 60-70ug/uL.
Thank you,

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Answer

Thank you for your reply and for confirming that information.

I've issued credit note # *** for this order. The full purchase price will be refunded back to your credit card pending approval of our accounting department. If you have any questions about this refund, please contact mailto:us.credits@abcam.com and refer to the credit note #.

I am sorry that this antibody did not work well, and if you have any questions or need anything else, please don't hesitate to ask.

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Question

Hello,
After several weeks of western blotting using various dilution factors of the Abcam anti-BNip3L (rabbit, #ab8399) antibody in human breast tumor cell samples, mouse neuron cell samples, and Alzheimer's disease cybrid cell samples, we are unsatisfied with this antibody's performance.
I have attached a PowerPoint with pictures displaying our concerns. In all samples tested, many non-specific bands were present during exposure in addition to the blots becoming VERY dirty. This is indeed caused by the BNip3L antibody since stripping and reprobing with Beta-actin and/or COX IV produces clean blots upon exposure.
Due to the unsatisfactory performance of anti-BNip3L (ab8399), we would like a refund.
Thank you,

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Answer

Thank you for contacting us and letting us know about the trouble with ab8399. I am sorry to see that the results were so disappointing.

I will be happy to issue a refund for your purchase. Please confirm your order information and I will process the refund.

The slides and information that you sent will be very helpful for our investigation into the problem. Could you also let me know the quantity of protein that was loaded per lane of the gel? I know that you've probably tried a few different loadings, so just anapproximaterange would be helpful.

I look forward to hearing from you and resolving this promptly. Please let me know if you have any questions or if there is anything else that we can do for you.

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Question

Could you help me figure out what the exact molecular weight for this protein is on a western blot with human tissue?
I used a 1:1000 dilution of anti-BNip3L on Human cybrid cell samples (neuroblastoma cell line with Non-Alzheimer's, Middle-staged Alzheimer's, and Late-stage Alzheimer's disease mitochondria). Three bands appeared on the blot following exposure.
After consulting numerous company information on google, it seems that the molecular weight of Nix/BNip3L should be 40kD. However, I do not believe that any of the bands are at 40 kD. Rather, two bands are below 37kD and one band is very close to 50kD. I believe the middle band nearest 37 kD should be the Nix protein band, but I would still like confirmation.

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Answer

Thank you for contacting Abcam.

Based on the information we have on this antibody, it is not uncommon for this antibody to recognize more than one band, but I would agree with your assessment that the band closet to 37kDa is the correct size band.

If you feel that the antibody is not functioning correctly and that it is not recognizing the correct band, then under our Abpromise, which is good for 6 months, we guarantee this antibody to work in western blot on human samples. Therefore if it does not work then we will either replace or refund the cost of this antibody.

Please let me know if there is anything else I can help you with.

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Question

Dear technical support team:
This customer has purchased ab8399 (Anti-BNIP3L antibody) and has conducted the wb with mouse sample. The results show weak signal, 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 data in this letter and her experiment step as follow:
1. Order details:
Batch number: XXXXX
Po: XXXXX
Abcam product code: ab8399
Antibody storage conditions (temperature/reconstitution etc) -20℃
2. Please describe the problem (high background, wrong band size, more bands, no band etc).
High background and weak signal
3. On what material are you testing the antibody in WB?
· Species: Mouse
· What’s cell line or tissue：liver
· Cell extract or Nuclear extract: cell extract
· Purified protein or Recombinant protein: Purified protein
3. The lysate
How much protein was loaded: 100 ug
What lysis buffer was used: EBC lysis buffer
What protease inhibitors were used: protease inhibitor (sigma)
What loading buffer was used: 4X loading dye
Phosphatase inhibitors ：no
Did you heat the samples: temperature and time:100℃ 10 min
4. Electrophoresis/Gel conditions/ Transfer conditions
Reducing or non reducing gel:
Reducing agent:
Gel percentage : 13％
Transfer conditions: (Type of membrane, Protein transfer verified): PVDF 100V 90 min
5. Blocking conditions
Buffer:TBST
Blocking agent: milk, BSA, serum, what percentage: 5％ milk
Incubation time:1 hr ℃
Incubation temperature: RT
6. Primary Antibody
Species: Rabbit
Reacts against: human and mouse
· At what dilution(s) have you tested this antibody:1:1000
· What dilution buffer was used: 5％ milk in TBST
· Incubation time: overnight
· Incubation temperature: 4℃
· What washing steps were done: four time , 10 min/each
7. Secondary Antibody
Species:Goat
Reacts against:Rabbit
At what dilution(s) have you tested this antibody: 1:5000
Incubation time: 1 hr RT
Wash steps: 15 min once, 10 min once, 5 min five times
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. NO
10. Optimization attempts
· How many times have you tried the Western? three
· Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no
· Do you obtain the same results every time e.g. are background bands always in the same place? yes
· What steps have you altered? We changed from PVDF to NC membrane last time
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 our products.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab8399.

-I would recommend the use of a positive control, as it is a good indicator of the antibody performance. Adipose tissue lysate is a good alternative.

-I would also recommend loading less protein sample per lane in order to reduce background. 20ug per lane could be a good starting point.

-Try to optimise the lysis process by using RIPA buffer which has a high denaturing level. I will also recommend adding phosphatase inhibitors.

-BNIP3L is a phosphorilated protein. Milk is not recommended as a blocking agent when phospho-proteins, as the casein could break the phosphorilated positions. Instead, I would recommend using 5% BSA for 2 hours at RT.

I hope this suggestions help improve the results from the antibody. Please do not hesitate to contact us for further assistance.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-BNIP3L/NIX antibody

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Foreskin Fibroblasts)
Permeabilization
Yes - 0.1% Triton X-100
Specification
Foreskin Fibroblasts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
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Dr. Sharon Sneddon

Verified customer

投稿 Nov 30 2011

Question

Can I also detect rat BNIP 3 with this antibody? Many thanks,

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Answer

Thank you for your interest in this antibody. We have only evaluated this antibody in human tissue and cell lines. However, it is possible for this antibody to also recognize BNIP 3 in rat.

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Question

Recently we purchased your rabbit anti-Nix (BNIP3L) antibody. We tried it on a variety of lysates including k562 lysates. Bad antibody. Very expoense, no specific signal and alot of backgroud. What can we do about this?

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Answer

I'm sorry that you are having problems with Rabbit polyclonal to Bnip3L. Indeed, K562 lystates are recommended as a positive control where a band of 40kDa corresponding to a monomer of Bnip3L is observed -- (as per reference 4 on the datasheet Oncogene (1999) 18:4523-9). I do not think that there is a problem with this antibody, but rather the WB conditions must be optimised to decrease background. I will need more information about your protocol and results in order to assist you. At what dilution(s) have you tested the antibody in WB, what are your blocking and incubation steps and how many bands do you observe on your blot? I look forward to hearing from you. Kind regards, Danielle N. Miller PhD Head of Customer Services

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Question

What is the price of this antibody and do you know whether it works for immunofluorescence studies. Best Brent

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Answer

So far, this antibody has only been tested in WB. Prices for 100 µg are as follows: GBP 195.00, USD 293.00, EUR 332.00 ( you may obtain this information by clicking on the prices and ordering tab - just to the right of the datasheet)

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