概述

  • 产品名称
    Anti-GST抗体[3G10/1B3]
    参阅全部 GST 一抗
  • 描述
    小鼠单克隆抗体[3G10/1B3] to GST
  • 宿主
    Mouse
  • 特异性
    Recognizes Glutathione S-Transferase (GST), a 26-kDa protein encoded by the parasitic helminth Schistosoma japonicum.
  • 经测试应用
    适用于: WB, IPmore details
  • 种属反应性
    与反应: Species independent
  • 免疫原

    Recombinant full length protein within Schistosoma japonicum GST aa 1-218. The exact sequence is proprietary. This is full length GST fusion protein of 425 aa (length of the fusion protein, not GST protein alone). The GST protein is the same GST protein on the pGEX vector ( Schistosoma japonicum).
    Database link: P08515

  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 25th May 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

性能

应用

Our Abpromise guarantee covers the use of ab92 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 0.5 - 2 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 26 kDa).
IP Use at an assay dependent dilution.

靶标

  • 相关性
    GST (Glutathione S-Transferase) is a 26kDa protein encoded by the parasitic helminth Schistosoma japonicum and widely used in the pGEX family of GST plasmid expression vectors as a fusion protein with foreign proteins.
  • 别名
    • Glutathione S Transferase antibody
    • Glutathione S-transferase class-mu 26 kDa isozyme antibody
    • GST antibody
    • Sj26 antigen antibody
    • SjGST antibody
    see all

图片

  • All lanes : Anti-GST antibody [3G10/1B3] (ab92) at 1/5000 dilution

    Lane 1 : Non-induced GST-fusion protein expressed in E.coli
    Lane 2 : IPTG induced GST-fusion protein expressed in E.coli

    Lysates/proteins at 5 µl per lane.

    Predicted band size: 26 kDa



    10% SDS-PAGE

  • All lanes : Anti-GST antibody [3G10/1B3] (ab92)

    Lane 1 : Total lysate
    Lane 2 : Cytoplasmic fraction
    Lane 3 : Membrane fraction
    Lane 4 : Nuclear fraction

    Predicted band size: 26 kDa



    Detection of human GST in cytoplasmic fraction by anti-GST 3G10 monoclonal antibody (ab92) in western blot experiment. p84 is also shown indicating the nuclear fraction using anti-p84 monoclonal antibody (ab487).

文献

This product has been referenced in:
See all 15 Publications for this product

客户评价及客户问答

1-8 of 8 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
HEK293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

Abcam user community

Verified customer

提交于 May 11 2018

Application
Western blot
Sample
Human Recombinant protein (Recombinant GST protein)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
0.2 µg
Specification
Recombinant GST protein
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Jan 06 2017

Answer

Thank you for your enquiry and your interest in our products.

I can confirm that we sell this antibody in liquid format, the unit size is 100 µg the concentration is 1 mg/ml.

If you need any further assistance in the future, please do not hesitate to contact me

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Answer

I am very glad to hear that our anti-Beta Actin loading control has worked so well for you.

In regards to a loading control for your current needs, Beta Actin would only be present in serum if shearing of cells had occurred during collection so this would not make a viable control. I have come across data stating that tranferrin and alpha anti-Trypsin are usually expressed at levels ˜2.5mg/ml and ˜1.3mg/ml respectively. However, as these may vary in samples, it may be best to add a purified peptide such a GST to your sample and stain against that.

We do offer antibodies against Transferrin, alpha anti-Trypsin and GST as well as purified GST peptides.

GST (ab70456):https://www.abcam.com/GST-protein-ab70456.html

anti-GST antibody (ab92):https://www.abcam.com/GST-antibody-3G10-1B3-ab92.html

anti-alpha-anti-Trypsin antibody (ab7633):https://www.abcam.com/alpha-1-Antitrypsin-antibody-ab7633.html

anti-Tranferrin antibody (ab1223):https://www.abcam.com/Transferrin-antibody-ab1223.html



I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Lysate from Insect Cells Expressing RGS9-GST fusio)
Loading amount
100 µg
Specification
Lysate from Insect Cells Expressing RGS9-GST fusio
Gel Running Conditions
Reduced Denaturing (10% Tris Glycine)
Blocking step
1:1 PBS/Licor Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C

Dr. Chris Tracy

Verified customer

提交于 Sep 20 2011

Question

BATCH NUMBER 10PX402 ORDER NUMBER 598807 DESCRIPTION OF THE PROBLEM smudged/fuzzy band(s)? SAMPLE Mouse liver samples PRIMARY ANTIBODY Mouse anti-GST (abcam #ab92) was used at a dilution of 1:1000 (3uL in 3mL of PBS/0.05% Tween20) This incubated with rotation for 2 hour at RT. The membrane was given a quick wash and then washed 4 x 5 min. with the PBS/Tween. DETECTION METHOD Western Blue Stabalized Substrate for AP (Promega #S3841) POSITIVE AND NEGATIVE CONTROLS USED nothing ANTIBODY STORAGE CONDITIONS Aliquoted with current tube stored at -20 degC and the rest stored at -80 degC. SAMPLE PREPARATION Livers were sonicated in NETT buffer pH 7.5 with Protease Inhibitor Cocktail Set III (EMD Biosciences #539134), adding the PI according to the weight of the livers. They were then diluted 1:10 with additional NETT buffer and 1:100 PI. It is quantitated and 20-50ug are used per lane. A 4x loading buffer is adding and the sample is heated at 95 degC for 5-10 min. AMOUNT OF PROTEIN LOADED The most recent was ~50 ug because we were having problems detecting lower levels of protein. ELECTROPHORESIS/GEL CONDITIONS 12% Polyacrylamide gel with stacking gel. These were run in a Tris-Glycine-SDS electrophoresis buffer for ~4 hours. It was run at 60V through the stacking gel and then 120-150V through the running gel. TRANSFER AND BLOCKING CONDITIONS They were transferred using a Tris-Glycine-Methanol buffer for 1 hour at 100V. They were then blocked overnight at 4 degC with 5% dry non-fat milk in PBS/0.2% Tween20/0.02%Na-azide. The membranes were subsequently rinsed with PBS/0.05%Tween20 before the primary antibody. SECONDARY ANTIBODY Goat anti-mouse AP (Promega #s3721) was used at a dilution of 1:7500 in PBS/0.05%Tween20). This incubated with rotation for 1 hour at RT (this Ab is known to work with other primary antibodies). The membrane was quickly washed and then 4 x 5 min. washes with the PBS/Tween. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? The first attempt was a dilution of the primary antibody at 1:500, the second was 1:2000, and this 3rd was 1:1000. The blot is always fuzzy. The primary has been left overnight at room temperature, overnight at 4 degC and 2 hours at room temp, respectively. ADDITIONAL NOTES I'm attaching an image of the blot. The bottom half is the portion stained for GST. The band(s)? is in the correct location, but it is so unclear that it is hard to tell if there are multiple bands and also difficult to quantitate because of this. The top section is not the top of this blot, but the top was stained with a different antibody and it had bands similar to this... so it is possible to stain with other antibodies and get nice bands.

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Answer

I'm sorry to hear you are experiencing problems with ab92. I have been able to source the images from the laboratory and have added them to the datasheet, interestingly the band we get with a HeLa whole cell lysate is also large ("fat") in a similar way to your blot so it may be that the protein's migration properties give this pattern. I enclose below other possible reasons for the problem which accentuate the band fuzziness: 1) too little Tween20 in the buffer: I would recommend adding more Tween20 to the antibody dilution buffer/washing buffer (0.1%v/v) and using a Tris based buffer rather than PBS as Tris gives sharper bands. 2) the percentage of SDS/methanol in the migration buffer and transfer buffer respectively can be a problem (SDS percentage should be around 1%w/v and should be fresh SDS, methanol percentage should be 20%v/v). The stacking gel composition can give problems with small proteins and resolution of small proteins often does not give as sharp a band as large proteins. 3) the blocking buffer can also sometimes give problems to the antibody and prevent it from binding correctly. I suggest trying 1hr milk 5% and incubation of the antibody in TBST only and also try 5%BSA the same way. 3) the lysis buffer (NETT buffer) may not be the most suitable extraction buffer for the protein. We do not have experience of this buffer and I would recommend trying a 20 mM Tris-HCl, pH 7.5 1 mM EGTA buffer or a NP40/RIPA buffer (50mM Tris HCl pH 8, 150 mM NaCl, 1 % NP-40, 0.5 % sodium Deoxycholate, 0.1 % SDS). 4) Finally, we have had good results with even more dilute antibody (0.5ug/ml equivalent to 1:2000) therefore you may find that by diluting the antibody more (and with the change in blocking buffer) and incubating at 4C overnight as you have previously done this will improve the signal (even if you need to expose the blot longer). I hope these suggestions will help you, please do not hesitate to contact me if you require further assistance,

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Answer

Thanks again for your email. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on a number of rewards. Regarding a catalogue, we don't have a traditional "paper" catalogue as all of our items are listed online on our website www.abcam.com Please let me know if you have any additional questions.

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Answer

Thank you for enquiry. I have checked with the originator of this antibody and there was a mistake regarding the immunogen on the datasheet. This antibody was generated as a bi-product during a fusion. The original immunogen was a GST fusion protein, with the total fusion protein being 425 amino acids. The actual GST portion is 232 amino acids: MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLIEGRGIPGNSS Unfortunately I am unable to share what the protein fragment was that was fused to GST during the generation of this particular clone due to confidentiality reasons. I can say is that it was a small fragment of a very high molecular weight protein. I have updated the datasheet to reflect this information and apologize for the inconvenience. If you have any additional questions, please contact us again.

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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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