PRODUCT CODE: ET1705-87

GCLM Recombinant Rabbit Monoclonal Antibody [JM93-61] (ET1705-87)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of GCLM on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-87, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A431 cell lysate<br />
Lane 2: PC-12 cell lysate<br />
Lane 3: NIH/3T3 cell lysate
  • Western blot analysis of GCLM on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-87, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A431 cell lysate<br />
Lane 2: PC-12 cell lysate<br />
Lane 3: NIH/3T3 cell lysate
  • ICC staining of GCLM in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of GCLM in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of GCLM in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-GCLM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-87, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-GCLM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-87, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue using anti-GCLM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-87, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of GCLM was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-87, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of GCLM on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-87, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A431 cell lysate
Lane 2: PC-12 cell lysate
Lane 3: NIH/3T3 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

GCLM Recombinant Rabbit Monoclonal Antibody [JM93-61] (ET1705-87)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

A431, PC-12, NIH-3T3, Hela, A549,human colon cancer tissue, human pancreas tissue, mouse small intestine tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JM93-61

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:100

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

GCLM

SYNONYMS

Gamma ECS regulatory subunit antibody; Gamma-ECS regulatory subunit antibody; Gamma-glutamylcysteine synthetase regulatory subunit antibody; GCLM antibody; GCS light chain antibody; GLCLR antibody; Glutamate cysteine ligase regulatory subunit antibody; Glutamate--cysteine ligase modifier subunit antibody; Glutamate--cysteine ligase regulatory subunit antibody; GSC light chain antibody; GSH0_HUMAN antibody

SEQUENCE SIMILARITIES

Belongs to the aldo/keto reductase family. Glutamate--cysteine ligase light chain subfamily.

TISSUE SPECIFICITY

In all tissues examined. Highest levels in skeletal muscle.

SUBCELLULAR LOCATION

Cytosol.

FUNCTION

Gamma-glutamylcysteine synthetase (γ-GCS) is the rate limiting enzyme for glutathione (L-gamma-glutamyl-L-cysteinylglycine, GSH) synthesis. GSH is ubiquitous in mammalian cells as a vital intra- and extracellular protective antioxidant. γ-GCS is a heterodimer of a heavy catalytic subunit and a light regulatory subunit that is responsive to inflammation, phenolic antioxidants, heat shock, oxidants and cytokines. The human gamma-GCS gene encoding the 367 amino acid catalytic subunit maps to chromosome 6p12. The human γ-GCS gene encoding the regulatory subunit maps to chromosome 1p22-p21. The two subunits of γ-GCS form a heterodimeric zinc metalloprotein that gains activity through formation of a reversible disulfide bond.