PRODUCT CODE: ET1611-5

Glutaminase Recombinant Rabbit Monoclonal Antibody [SN68-09] (ET1611-5)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of Glutaminase 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 (ET1611-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: 293 cell lysate
  • Western blot analysis of Glutaminase 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 (ET1611-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: 293 cell lysate
  • ICC staining of Glutaminase 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 (ET1611-5, 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 Glutaminase in HepG2 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 (ET1611-5, 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 tonsil tissue using anti-Glutaminase 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 (ET1611-5, 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 kidney tissue using anti-Glutaminase 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 (ET1611-5, 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.
Western blot analysis of Glutaminase 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 (ET1611-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: 293 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Glutaminase Recombinant Rabbit Monoclonal Antibody [SN68-09] (ET1611-5)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

293, Hela, HepG2, human tonsil tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SN68-09

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.

MOLECULAR WEIGHT

65 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Glutaminase

SYNONYMS

AAD20 antibody; DKFZp686O15119 antibody; FLJ10358 antibody; GAC antibody; GAM antibody; GLS antibody; GLS1 antibody; GLSK_HUMAN antibody; Glutaminase C antibody; Glutaminase kidney isoform antibody; Glutaminase phosphate activated antibody; K-glutaminase antibody; KGA antibody; KIAA0838 antibody; L-glutamine amidohydrolase antibody; mitochondrial antibody

SEQUENCE SIMILARITIES

Belongs to the glutaminase family.

TISSUE SPECIFICITY

Isoform 1 and isoform 3 are detected in brain cortex. Isoform 3 is highly expressed in astrocytoma, ganglioglioma and ependymoma. Isoform 1 is highly expressed in brain and kidney, but not detected in liver. Isoform 3 is highly expressed in heart and pancreas, detected at lower levels in placenta, lung, pancreas and kidney, but is not detected in liver. Isoform 2 is expressed in cardiac and skeletal muscle.

POST-TRANSLATIONAL MODIFICATION

Synthesized as a 74-kDa cytosolic precursor which is proteolytically processed by the mitochondrial-processing peptidase (MPP) via a 72-kDa intermediate to yield the mature mitochondrial 68- and 65-kDa subunits.

SUBCELLULAR LOCATION

Cytoplasm, Mitochondrion.

FUNCTION

Glutamine is an important molecule involved in several cellular functions, including nitrogen and carbon transport, hepatic urea synthesis, renal ammoniagenesis, and gluconeogenesis. Glutamine is catabolized by either the liver-type (LGA) or kidney-type (KGA) glutaminase. KGA is mitochondrial specific protein whose expression in kidney is increased during metabolic acidosis. This process is mediated by an 8-base AU-sequence in KGA that functions as a pH-response element. The human KGA gene maps to chromosome 2, and produces three isoforms, designated KGA, GAC, and GAM, by alternative splicing. KGA is synthesized as a cytosolic protein that is transported to the mitochondria as an intermediate protein, and is further cleaved into the KGA isoform and the GAC isoform. The processing of the GAM isoform is unclear. The KGA isoform is abundant in brain and kidney, while the GAC isoform is principally expressed in cardiac muscle and pancreas. The GAM isoform is solely expressed in cardiac and skeletal muscle.