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  1. Link

    mouse-brain-tissue-lysate-total-protein-14-days-ab7189.pdf

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Kits/ Lysates/ Other Lysates Tissue Lysates Mouse Nervous System
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Mouse brain tissue lysate - total protein (14 days) (ab7189)

  • Datasheet
  • SDS
Submit a review Q&A (5)

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Overview

  • Product name

    Mouse brain tissue lysate - total protein (14 days)
    See all Brain lysates
  • General notes

    Source: Outbred NIH-Swiss Albino mice.

    The mouse brain tissue was collected from fourteen (14) day-old mice. The lysate was prepared by homogenization in modified RIPA buffer (50 mM Tris -HCl, pH 7.4, 1% Triton X-100, 0.2% sodium deoxycholate, 0.2% sodium dodecylsulfate (SDS), 1 mM sodium ethylenediaminetetraacetate, 1 mM phenylmethylsulfonyl flouride, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The tissue lysate was boiled for 5 min in 1 x SDS sample buffer (0.045 M Tris-HCl pH 6.8, 10% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue), containing 0.05 M DTT.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Mycoplasma free

    Yes
  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Constituent: 100% SDS Sample Buffer
  • Concentration information loading...
  • Research areas

    • Kits/ Lysates/ Other
    • Lysates
    • Tissue Lysates
    • Mouse
    • Nervous System

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab7189 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
WB
Use at an assay dependent dilution. 10 µg to 20 µg per lane is recommended for mini gel.
Notes
WB
Use at an assay dependent dilution. 10 µg to 20 µg per lane is recommended for mini gel.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab7189? Please let us know so that we can cite the reference in this datasheet.

ab7189 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Question

nach längerer Suche kam ich nicht zu dem gleichen Ergebnisse der Enzym Größe. Wie kommen sie auf 138/148 kDa?

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Abcam community

Verified customer

Asked on Aug 30 2012

Answer

Wie besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 1155271 und sollte morgen bei Ihnen ankommen.

Das Molekulargewicht kann man folgendermaßen errechnen: Öffnen Sie die SwissProt Link auf unserem Datenblatt für das Maus Protein (http://www.uniprot.org/uniprot/P09470), und rollen Sie die Seite einmal bis zum Abschnitt "Sequence annotation (Features)" herunter. Dort finden Sie die Angaben zum "Molecule processing" ,in welchen die Länge und Positionen der einzelnen Protein Ketten beschrieben werde. Wenn Sie die jeweilige Position öffnen, gelangen Sie zur FASTA Sequenz ( in dem Kasten oben links).

Desweiteren oeffnen Sie den Molekular Gewicht/ pI Rechner unter dem folgenden Link http://web.expasy.org/compute_pi/

In diesen fügen Sie die FASTA Sequenz ein, und lassen das Gewicht berechnen.

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

Read More

Abcam Scientific Support

Answered on Aug 30 2012

Question

sehr gerne schicken Sie mir die positiv Kontrolle. Wann könnte ich damit rechnen?

Read More

Abcam community

Verified customer

Asked on Aug 29 2012

Answer

Vielen Dank fürIhre Anfrage.

Da das Lysat bei uns vorrätig ist, kann es am nähsten Tag geliefert werden. Wenn Sie mir also die Lieferadresse mitteilen, kann ich den Versandt gleich abwickeln. Es sollte übrigens kein Problem sein, es direkt an Sie ohne den Umweg über die Apotheke zu schicken, da es sich ja um einen kostenlosen Ersatz handelt.

Ich freue mich auf Ihre Antwort.

Read More

Abcam Scientific Support

Answered on Aug 29 2012

Question

Danke für die schnelle Bearbeitung. Anbei das troubleshooting und drei ausgewählte blots meiner inkubierten Membran.
Vielen lieben Dank im Vorraus,
Marion Bisha
1) Abcam product code ab11737 and ab75762

2) Abcam order reference number or product batch number
3) Description of the problem : wrong so signals
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates, bEND 3 (brain endothelial cells)
Lysis buffer : Rippa buffer 1ml, PIC 40µl, NaF 10 µl, NaP2O 10 µl, BGP 10 µl, NaV 5 µl
Boiling for ≥5 min? no 5 min 95°C
Protein loaded ug/lane 40 Mg / lane
Positive control ACE rabbit
Negative control
5) Percentage of gel: 10%
Type of membrane : PVDF
Protein transfer verified
Blocking agent and concentration; Odyssey Blocking buffer 50% / PBS 50%
Blocking time : 1 hour
Blocking temperature room temperature
6) Primary antibody:
Concentration or dilution : ab11737= 1:100 and 1:50 and 1:20 ab75762 = 1:1000
Diluent buffer : Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween
Incubation time: over night or 4 hours
Incubation temperature: over night inn 4 °C and 4 hours at room temperature
7) Secondary antibody: ab11737: anti mouse Odyssey antibody
Ab75762 anti rabbit Odyssey antibody
Species:
Reacts against:
Concentration or dilution 1:10000
Diluent buffer : Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween
Incubation time: 1 hour
Incubation temperature: room temperature
Fluorochrome or enzyme conjugate:
8) Washing after primary and secondary antibodies:
Buffer: between the antibodies incubation 4 x 5 min with Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween and after the incubation with the second antibody 2 x 5 min with Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween and 2 x 5 min with Odyssey Blocking buffer 50% / PBS 50%
Number of washes
9)Detection method
10) How many times have you run this staining? 5 x
Do you obtain the same results every time? Jes
What steps have you altered to try and optimize the use of this antibody?
Different concentration of the antibodies and different incubation times
Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Read More

Abcam community

Verified customer

Asked on Aug 28 2012

Answer

Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.



Ich möchte folgende Bemerkungen zu Ihrem Protokoll machen:

1. Ich denke, Sie detektieren das richtige Protein, da zwei so unterschiedliche Antikörper das gleiche Signal zeigen. Dies kann am veränderten Stoffwechsel der Zellen liegen, die Sie verwenden ( schließlich handelt es sich hier laut ATCC um ein Endothelioma).


2. Schauen Sie sich bitte einmal das Dokument an, welches ich an diese Email gehängt habe: Dieses Dokument zeigt sehr schön, wie das Gelsystem das Laufverhalten beeinflußen kann.

3. Eine Abweichung von bis zu 10% von der erwarteten Groesse kann durchaus als normal angesehen werden.

4. Es gibt zwei Formen dieses Proteins, die lösliche und die zelluläre Form. Die lösliche From ist ca 10 kDa leichter als die zellulare (138 kDa vs 148 kDA).

5. Da Sie leider mit einem System arbeiten (Odyssey), ist es schwierig Veränderungsvorschläge zu Ihrem Blot zu machen. Es würde mich allerdings interessieren, ob Sie via Chemolumineszenz oder Fluoreszenz detektieren.

6. Um auszuschließen, dass Sie das falsche Protein detektieren, möchte ich Ihnen ein Maushirn Lysat als Positiv- bzw Vergleichsprobe kostenlos zusenden. Bitte lassen Sie mich wissen, ob Sie damit einverstanden sind.

Read More

Abcam Scientific Support

Answered on Aug 28 2012

Question

Do you guys have a positive control for Anti-Cdk2 antibody (ab6538)? I would like to purchase this antibody with a tested positive control. Thanks! sincerely,  

Read More

Abcam community

Verified customer

Asked on Sep 12 2011

Answer

Thank you for contacting us. I presume you are looking for positive control used in western blot. Our customers have used this ab successfully with mouse brain tissue and 293 cells lysates. Please check the Abreviews. The Mouse brain lysates can be found with catalogue numbers are; ab7189, ab7190 ab30151 and Human 293 cell lysates with catalogue number ab7902. Please also click the following link to see the tissues which express CDK2 target, lysates of these can be used as positive control. http://www.proteinatlas.org/ENSG00000123374/normal http://www.proteinatlas.org/ENSG00000123374/cancer http://www.proteinatlas.org/ENSG00000123374/cell We have the tissue lysates in catalogue. Please type "lysates" in the product search box provide on the Abcam homepage and system will give you sorted products as per your requirements. 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 Sep 12 2011

Question

How many mouse brains does it take to produce the 100ug of ready to load lysate? thanks.

Read More

Abcam community

Verified customer

Asked on Feb 14 2008

Answer

Thank you for your enquiry. We do not produce 100ug at a time, but many milligrams from multiple mouse brains. The exact number can vary with each batch. The brain has a lot of protein, so it may only take one mouse brain to get 100ug. I hope this information will be helpful. 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 Feb 14 2008

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