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GSTM1 Polyclonal Antibody

Uniprot : P10649
  • Cat.No.:E-AB-40289

  • Host: Rabbit
  • Reactivity: H,M,R
  • Applications: WB,IHC,IF

To Purchase E-AB-40289

Size:
  • 20μL
  • 60μL
  • 120μL
  • 200μL
Price: $65
Qty:

Test Application

  • Verified Samples

    Reactivity Application
    Human WB
    (Hela,Jurkat,)

    Western Blot analysis of Hela cells using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    IHC
    (liver cancer,kidney,)

    Immunohistochemistry of paraffin-embedded Human liver cancer using GSTM1 Polyclonal Antibody at dilution of 1:100.

    Immunohistochemistry of paraffin-embedded Human kidney using GSTM1 Polyclonal Antibody at dilution of 1:100.

    IF
    (Hela,Hela,HepG2,)

    Immunofluorescence analysis of Hela cells using GSTM1 Polyclonal Antibody at dilution of 1:100.

    Immunofluorescence analysis of Hela cells using GSTM1 Polyclonal Antibody at dilution of 1:100.

    Immunofluorescence analysis of HepG2 cells using GSTM1 Polyclonal Antibody at dilution of 1:100.

    Rat WB
    (kidney,heart,)

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Mouse WB
    (liver,heart,NIH/3T3,kidney,)

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

    Western Blot analysis of Mouse liver, Mouse heart, NIH/3T3 cells, Jurkat cells, Rat kidney, Rat heart and Mouse kidney using GSTM1 Polyclonal Antibody at dilution of 1:500.

  • Dilution

    WB 1:500-1:6000 IHC 1:100-1:200 IF 1:100-1:200

  • Western Blot Operation Guide

    In order to facilitate the operation and ensure the accuracy of WB results, the Western Blot Detection kit (Cat# E-IR-R304) is now available, containing the reagents which are needed from sample preparation to result detection. Please order the appropriate kit according to your specific needs.

    Separating gel15%
    Blocking1.5 h
    Primary Antibody1:500-1:4000
    Secondary Antibody1:5000
    Click Here for More Details .. More ↓

Preparation of protein samples

1.Protein extraction

1)For tissue sample
a. Take the samples, wash the tissue thoroughly with pre-cooled PBS (0.01 M, pH=7.4)(Cat# E-BC-R187) to remove the surface blood and internal debris.
b. Weigh and smash the tissue, add an appropriate ratio of RIPA Lysis Buffer (Cat# E-BC-R327)(add 10 μL PMSF and 10 μL Na3VO4 to each 1 mL RIPA Lysis) and homogenizely lyse the tissue.
It is recommended to homogenize according to the ratio of tissue weight: RIPA volume = 3:10. For example, add 1 mL RIPA Lysis Buffer to 0.3 g tissue sample, the specific volume can be adjusted according to experimental requirements.
c. Shake and lyse on the ice for 30 min after homogenization. And then sonicate the sample for 1 min (under ice water bath conditions) with 2 s’ sonication and 2 s’ intervals to make cells fully lysis and reduce the viscosity of sample.
d. Centrifuge at 12,000 rpm for 10 min at 4℃.
e. Take the supernatant and measure the protein concentration mentioned in step2.

2)For cell sample
a. Collect the cells, wash them thoroughly with pre-cooled PBS (0.01 M, pH=7.4) to remove the medium off (it is generally recommended to wash 3 times).
b. Add an appropriate ratio of RIPA Lysate Buffer (10 μL PMSF and 10 μL Na3VO4 in each 1 mL RIPA Lysis) and lyse on the ice for 30 min.
It is recommended to add 0.1 mL of RIPA Lysis Buffer to each well of a 6-well plates (the protein content in different cells may vary, and the volume of the lysate added can be appropriately adjusted).
c. Sonicate the sample for 1 min (under ice water bath conditions) with 2 s’ sonication and 2 s’ intervals to make cells fully lyse and reduce viscosity of sample.
d. Centrifuge at 12,000 rpm for 10 min at 4℃.
e. Take the supernatant and measure the protein concentration mentioned in step2.

2.Measurement of protein concentration
By the BCA method (see the Total Protein Colorimetric Assay Kit (Cat# E-BC-K318) instructions).

3.Boiling the samples
Adjust the protein concentration with PBS Buffer. Add 5 × SDS Loading Buffer (Cat# E-BC-R288) with the ratio of the protein sample: 5 × SDS Loading Buffer = 4:1 and boil the mixture for 10 min. Centrifuge at 12,000 rpm for 2 min and collect the supernatant. The denatured protein can be employed to Western Blot experiments or stored at -20℃ or -80℃.

Note: It is recommended that the total protein loading amount of test sample is about 50 μg in each well. Try to make the loading volume of each sample close to 10 μL.

Electrophoresis

1.According to the molecular weight of the target protein, prepare 15% separation gel. Add the test sample to each well, and add 5 μL of Pre-stained Protein Marker (Cat# E-BC-R273)to a reserved well in order to verify the target molecular weight and the extent of membrane transfer. Add Electrophoresis Buffer ( Cat# E-BC-R331) and start electrophoresis.

2.Electrophoresis at 80v when the samples are in stacking gel, then convert to 120v when the blue flow into the separating gel. Electrophoresis time is about 2-3 h till bromophenol blue reaches the bottom of the gel.

Transfer Membrane

1.Choose the PVDF Membrane (Cat# E-BC-R266) with a pore size of 0.45 μm according to the molecular weight of the target protein. Soak the PVDF Membrane in methanol for 1 min to activate it, and then soak the PVDF Membrane in the Transmembrane Buffer (Cat# E-BC-R333), the filter paper and fiber mat must be soaked in the Transmembrane Buffer for use too.

2.Follow manufacture instructions of Transfer System for wet, semi-dry, or dry transfer.

Incubation of antibodies

1.Soak the PVDF Membrane with TBST Buffer (Cat# E-BC-R335) containing 5% Skim Milk Powder as blocking buffer and block the membrane at room temperature for 1.5 h.

2.According to the recommended primary antibody dilution ratio, use the TBST Buffer containing 5% Skim Milk Powder to dilute the GSTM1 Antibody at 1:500-1:4000, soak the PVDF Membrane in the primary antibody working solution, incubate overnight at 4 ℃, and gently shake.

3.Wash the PVDF Membrane with TBST Buffer for 3 times, 15 min/time.

4.According to the recommended secondary antibody dilution ratio, use a TBST Buffer solution containing 2% Skim Milk Powder to dilute Goat Anti-Rabbit IgG (H+L) (peroxidase/HRP conjugated) (Cat# E-AB-1003) at 1:5000. Incubate at room temperature for 1 h on a shaker.

5.Wash the PVDF Membrane with TBST Buffer for 3 times, 15 min/time.

Detection

1.Mix A and B in the Excellent Chemiluminescent Substrate Detection kit (Cat# E-BC-R347) at the ratio of 1:1 as working solution.

2.Take out the PVDF Membrane from TBST Buffer and absorb the liquid with the filter paper. Pave the PVDF Membrane on the detection machine, add ECL working solution continuously on the PVDF Membrane, discharge the bubble and detect the result.

3.Adjust the contrast and the exposure time to get the best image.

Appendix

Product Details

Clonality Polyclonal
Isotype IgG
Concentration 0.54 mg/mL
Storage Store at -20℃. Avoid freeze / thaw cycles.
Buffer PBS with 0.05% Proclin300 and 50% glycerol, pH7.4.
Purification Method Antigen Affinity Purification
Research Areas Tags and Cell markers,Signal Transduction,Metabolism
Conjugation Unconjugated

Immunogen Details

Immunogen Recombinant Mouse Glutathione S-transferase Mu 1 protein(PKSM041382)
Abbre GSTM1
Synonyms GST class-mu 1, GST HB subunit 4, GST1, Gstm1, GSTM1a-1a, GTH4, GTM1, H-B, MU
Swissprot P10649
Gene ID 14862
Calculated MW 26 kDa
Observed MW 26 kDa
Cellular Localization Cytoplasm
Tissue Specificity Highest expression level in urinary bladder urothelium

Background

Cytosolic and membrane-bound forms of glutathione S-transferase are encoded by two distinct supergene families. At present, eight distinct classes of the soluble cytoplasmic mammalian glutathione S-transferases have been identified: alpha, kappa, mu, omega, pi, sigma, theta and zeta. This gene encodes a glutathione S-transferase that belongs to the mu class. The mu class of enzymes functions in the detoxification of electrophilic compounds, including carcinogens, therapeutic drugs, environmental toxins and products of oxidative stress, by conjugation with glutathione. The genes encoding the mu class of enzymes are organized in a gene cluster on chromosome 1p13.3 and are known to be highly polymorphic. These genetic variations can change an individual's susceptibility to carcinogens and toxins as well as affect the toxicity and efficacy of certain drugs. Null mutations of this class mu gene have been linked with an increase in a number of cancers, likely due to an increased susceptibility to environmental toxins and carcinogens. Multiple protein isoforms are encoded by transcript variants of this gene.

Reviews/Q&A

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  • Reviews
  • Q&A

Verified Customer

P*******gSubmitted [ Dec 31 2019 ]

  • Application:WB
  • Species:Mouse
  • Loading amount:10μg
  • Sample source:Mouse liver
  • Gel Running Conditions:Reduced, Denaturing, Other details:15%
  • Blocking:

    Blocking buffer:Milk

    Blocking concentration:5 %

    Blocking temperature:25℃

    Blocking time: 1hours 30minutes

  • Primary antibody:

    Dilution:1:2000

    Time: 18hours

  • Temperature:4℃

    Diluent:5% Milk

  • Secondary antibody:Use Elabscience secondary antibody
  • Dilution:1:5000
  • Detection:

    method:ECL

  • Description:Western Blot analysis of Mouse liver using GSTM1 Polyclonal Antibody at dilution of 1:2000.
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Verified Customer

B********gSubmitted [ Nov 23 2019 ]

  • Application:WB
  • Species:Human
  • Loading amount:10μg
  • Sample source:Hela
  • Gel Running Conditions:Reduced, Denaturing, Other details:15%
  • Blocking:

    Blocking buffer:Milk

    Blocking concentration:5 %

    Blocking temperature:25℃

    Blocking time: 1hours 30minutes

  • Primary antibody:

    Dilution:1:500

    Time: 17hours

  • Temperature:4℃

    Diluent:5% Milk

  • Secondary antibody:Use Elabscience secondary antibody
  • Dilution:1:5000
  • Detection:

    method:ECL

  • Description:Western Blot analysis of Hela cells using GSTM1 Polyclonal Antibody at dilution of 1:500.
Read More
Read Less

Verified Customer

s****lSubmitted [ Nov 07 2019 ]

  • Application:WB
  • Species:Mouse
  • Loading amount:50μg
  • Sample source:mouse heart,mouse kidney
  • Gel Running Conditions:Reduced, Denaturing, Other details:15
  • Blocking:

    Blocking buffer:Milk

    Blocking concentration:5 %

    Blocking temperature:25℃

    Blocking time: 1.5hours

  • Primary antibody:

    Dilution:1:500

    Time: overnhours

  • Temperature:4℃

    Diluent:5% skim milk

  • Secondary antibody:Use Elabscience secondary antibody E-AB-1003
  • Dilution:1:5000
  • Detection:

    method:ECL

  • Description:The result is pretty good.There is no other non-specific band.
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