Anti-beta Actin 抗体 - Loading Control (ab8229)

製品の概要

  • 製品名
    Anti-beta Actin antibody - Loading Control
    beta Actin 一次抗体 製品一覧
  • 製品の詳細
    Goat polyclonal to beta Actin - Loading Control
  • 由来種
    Goat
  • 特異性
    This antibody detects a single clean band at just above 40kD that corresponds to beta Actin. Please note this antibody is raised against the b-nonmuscle isoform and does not stain adult cardiac and skeletal muscles, except for traces due to contaminations of the sample with non-muscle cells, or if embryonic tissue is being used. The immunogen used for this product shares 77% homology with Gamma actin/actin cytoplasmic 2. Cross-reactivity with this protein has not been confirmed experimentally.
  • アプリケーション
    適用あり: WBmore details
  • 種交差性
    交差種: Mouse, Rat, Rabbit, Goat, Chicken, Cow, Dog, Human, Fish, Monkey, Chinese hamster
    交差が予測される動物種: a wide range of other species
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human beta Actin aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab13772)

  • ポジティブ・コントロール
    • WB: HeLa, NIH3T3, PC12 and CHOK1 whole cell lysates.
  • 特記事項

    This antibody has been designed for use as a loading control and is ideal for this purpose.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab8229 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 1/500. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772).

ターゲット情報

画像

  • All lanes : Anti-beta Actin antibody - Loading Control (ab8229) at 1/1000 dilution

    Lane 1 : HeLa whole cell (Human)
    Lane 2 : 3T3 cell (Mouse)
    Lane 3 : Rabbit Liver
    Lane 4 : MDCK cell (Dog)

    Lane 5 : EBTr cell (Cow)
    Lane 6 : SL-29 cell (Chicken)
    Lane 7 : CHO cell (Chinese Hamster)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Rabbit Anti-Goat IgG H&L (HRP) (ab6741) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 5 minutes


    Secondary antibody - rabbit anti-goat HRP (ab6741)

  • Lanes 1-4 : Anti-beta Actin antibody - Loading Control (ab8229) at 1 µg/ml (Blocked with 5% BSA.)
    Lanes 5-8 : Anti-beta Actin antibody - Loading Control (ab8229) at 1 µg/ml (Blocked with 3% Milk.)

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lanes 2 & 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lanes 3 & 7 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lanes 4 & 8 : CHO-K1 cell lysate Whole Cell Lysate
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa
    Additional bands at: 48 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin (lanes 1-4) or 3% milk (lanes 5-8) before being incubated with ab8229 overnight at 4°C. Antibody binding was detected using an anti-Goat antibody conjugated to HRP, and visualised using ECL development solution.  A non-specific band at 50-kDa is observed on western blots of some of our batches of ab8229. We have found that milk blocking is more effective than BSA blocking in eliminating this non-specific binding. 

プロトコール

参考文献

This product has been referenced in:
  • Tran KV  et al. Distinct adipocyte progenitor cells are associated with regional phenotypes of perivascular aortic fat in mice. Mol Metab 9:199-206 (2018). WB ; Mouse . Read more (PubMed: 29396370) »
  • Oropeza-Almazán Y  et al. Small Interfering RNA Targeting Mitochondrial Calcium Uniporter Improves Cardiomyocyte Cell Viability in Hypoxia/Reoxygenation Injury by Reducing Calcium Overload. Oxid Med Cell Longev 2017:5750897 (2017). Read more (PubMed: 28337252) »

See all 88 Publications for this product

レビューと Q&A

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Cow Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Dec 04 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cancer cell lines)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Colon cancer cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
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投稿 Jul 29 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
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投稿 May 07 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (neuronal)
Loading amount
30 µg
Specification
neuronal
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%
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Dr. Nicola Bright

Verified customer

投稿 Nov 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human C4-2 cells)
Loading amount
20 µg
Specification
human C4-2 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
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投稿 Nov 17 2011

Application
Western blot
Sample
Human Cell lysate - nuclear (PBMC)
Loading amount
20 µg
Specification
PBMC
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Oct 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Brain tissue)
Loading amount
25 µg
Specification
Brain tissue
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
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投稿 Oct 30 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Loading amount
5 µg
Specification
MEF
Gel Running Conditions
Reduced Denaturing (4-12% Nupage gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Mar 18 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Mixed Hippocampal Culture)
Loading amount
100 µg
Specification
Mixed Hippocampal Culture
Treatment
24hrs
Gel Running Conditions
Non-reduced Denaturing (10% Tris HCl)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
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投稿 Feb 01 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Loading amount
25 µg
Specification
HeLa cells
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Karppiah Muthumani

Verified customer

投稿 Aug 02 2007

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