Anti-beta Actin抗体- Loading Control (ab8229)

概述

  • 产品名称Anti-beta Actin抗体- Loading Control
    参阅全部 beta Actin 一抗
  • 描述
    山羊多克隆抗体to beta Actin - Loading Control
  • 特异性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.
  • 经测试应用适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Goat, Chicken, Cow, Dog, Human, Fish, Monkey, Chinese hamster
    预测可用于: 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)

  • 阳性对照
    • WB: HeLa, NIH3T3, PC12 and CHOK1 whole cell lysates. IHC-P: Human normal colon tissue. ICC/IF: HeLa cells.
  • 常规说明

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

性能

应用

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.

应用 Ab评论 说明
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB 1/500. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772).

靶标

Anti-beta Actin antibody - Loading Control 图像

  • 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
    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)

  • ICC/IF image of ab8229 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab8229, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal donkey serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • IHC image of beta Actin staining in Human normal colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8229, 10µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab6885), donkey anti-goat HRP (IgG - H&L; 1/250 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is taken from an identical assay without primary antibody.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    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. 

Anti-beta Actin antibody - Loading Control (ab8229)参考文献

This product has been referenced in:
  • Sun SH  et al. SIX1 coordinates with TGFß signals to induce epithelial-mesenchymal transition in cervical cancer. Oncol Lett 12:1271-1278 (2016). WB ; Human . Read more (PubMed: 27446426) »
  • Tamm C  et al. Fast and Efficient Transfection of Mouse Embryonic Stem Cells Using Non-Viral Reagents. Stem Cell Rev 12:584-591 (2016). Read more (PubMed: 27358240) »

See all 52 Publications for this product

Product Wall

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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Abcam user community

Verified customer

提交于 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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Abcam user community

Verified customer

提交于 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

提交于 Nov 28 2012

Thank you for your message. I am sorry to hear the free of charge replacement is also not providing satisfactory results and that this antibody is not working for you.

I appreciate the time you have spent on these experiments.I would suggest...

Read More

We have not tested ab8229 on swine samples, but based on the level of homology between the immunogen sequence and the swine sequence (96%), I would expect ab8229 to bind to swine samples, although we cannot guarantee it.

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

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提交于 Feb 01 2008

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