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  1. Link

    gst-antibody-3g101b3-ab92.pdf

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Tags & Cell Markers Fusion / Marker Proteins GST
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Anti-GST antibody [3G10/1B3] (ab92)

  • Datasheet
Reviews (3)Q&A (5)References (24)

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Western blot - Anti-GST antibody [3G10/1B3] (ab92)
  • Western blot - Anti-GST antibody [3G10/1B3] (ab92)

Key features and details

  • Mouse monoclonal [3G10/1B3] to GST
  • Suitable for: WB, IP
  • Reacts with: Species independent
  • Isotype: IgG1

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Overview

  • Product name

    Anti-GST antibody [3G10/1B3]
    See all GST primary antibodies
  • Description

    Mouse monoclonal [3G10/1B3] to GST
  • Host species

    Mouse
  • Specificity

    Recognizes Glutathione S-Transferase (GST), a 26-kDa protein encoded by the parasitic helminth Schistosoma japonicum.
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    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

  • General notes

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.40
    Constituent: 100% PBS

    w/o preservative
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    3G10/1B3
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GST
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Fusion/marker protein ELISA Kits

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Recombinant Protein

    • Recombinant S. japonicum GST protein (ab81793)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
WB (3)
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.
Notes
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.

Target

  • Relevance

    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.
  • Alternative names

    • Glutathione S Transferase antibody
    • Glutathione S-transferase class-mu 26 kDa isozyme antibody
    • GST antibody
    • Sj26 antigen antibody
    • SjGST antibody
    see all

Images

  • Western blot - Anti-GST antibody [3G10/1B3] (ab92)
    Western blot - Anti-GST antibody [3G10/1B3] (ab92)
    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

  • Western blot - Anti-GST antibody [3G10/1B3] (ab92)
    Western blot - Anti-GST antibody [3G10/1B3] (ab92)
    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).

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (24)

Publishing research using ab92? Please let us know so that we can cite the reference in this datasheet.

ab92 has been referenced in 24 publications.

  • Zhang X  et al. PTPN22 interacts with EB1 to regulate T-cell receptor signaling. FASEB J 34:8959-8974 (2020). PubMed: 32469452
  • Marquardt J  et al. The LKB1-like Kinase Elm1 Controls Septin Hourglass Assembly and Stability by Regulating Filament Pairing. Curr Biol 30:2386-2394.e4 (2020). PubMed: 32386534
  • Geymonat M  et al. Orderly assembly underpinning built-in asymmetry in the yeast centrosome duplication cycle requires cyclin-dependent kinase. Elife 9:N/A (2020). PubMed: 32851976
  • Pantier R  et al. TET1 Interacts Directly with NANOG via Independent Domains Containing Hydrophobic and Aromatic Residues. J Mol Biol 432:6075-6091 (2020). PubMed: 33058869
  • He Y  et al. The OsGSK2 Kinase Integrates Brassinosteroid and Jasmonic Acid Signaling by Interacting with OsJAZ4. Plant Cell 32:2806-2822 (2020). PubMed: 32586913
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-8 of 8 Abreviews or Q&A

Western blot abreview for Anti-GST antibody [3G10/1B3]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 11 2018

Western blot abreview for Anti-GST antibody [3G10/1B3]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 06 2017

Western blot abreview for Anti-GST antibody [3G10/1B3]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Chris Tracy

Verified customer

Submitted Sep 20 2011

Question

Inquiry: Dear Abcam team, I just want to double check that this antibody comes as a powder/lypholyzed sample that I can dilute to the concentration I want. https://www.abcam.com/GST-antibody-3G10-1B3-ab92.html I am inquiring because I require a 0.5mg/ml concentration of anti-GST for arraying on protein microarrays. Best regards,

Read More

Abcam community

Verified customer

Asked on Dec 13 2012

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

Read More

Abcam Scientific Support

Answered on Dec 13 2012

Question

Hi,   I am wondering what would you recommend me to use as a loading control for human serum sample? Your website only showed what you have available, but I want to know what I can use?  I have been your beta-actin for most of my experiment and it works great!

Read More

Abcam community

Verified customer

Asked on Apr 03 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 03 2012

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.

Read More

Abcam community

Verified customer

Asked on Oct 04 2005

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,

Read More

Abcam Scientific Support

Answered on Oct 05 2005

Question

Thanks for your fast reply. I wonder if you would like to reward me another free sample of this product or a catalog of your company? That would be more appreciated.

Read More

Abcam community

Verified customer

Asked on Feb 08 2005

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.

Read More

Abcam Scientific Support

Answered on Feb 08 2005

Question

on the datasheet, it's said this antibody was raise by targetting 1-425aa of human GST. I didn't find any HUMAN GST had 425aa, normally they were 200aa. I need to know the exact seqence of the antigen which you used to raise this antibody.

Read More

Abcam community

Verified customer

Asked on Feb 03 2005

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.

Read More

Abcam Scientific Support

Answered on Feb 08 2005

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