KGA/GAC Monoclonal antibody

• Featured Product
• KD/KO Validated

KGA/GAC Monoclonal Antibody for IF, IHC, WB, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, pig, mouse, rat

Applications

WB, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

6

CloneNo.

3A12A1

Reviews (1)

Cat no : 66265-1-Ig

Synonyms

AAD20, GAC, GLS, GLS1, glutaminase, K glutaminase, KGA, KIAA0838, L glutamine amidohydrolase

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Validation Data Gallery

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  WB analysis of NIH/3T3 using 66265-1-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 66265-1-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat liver using 66265-1-Ig

  rat liver tissue were subjected to SDS PAGE followed by western blot with 66265-1-Ig (GLS Antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of fetal human brain using 66265-1-Ig

  fetal human brain tissue were subjected to SDS PAGE followed by western blot with 66265-1-Ig (GLS Antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of U-251 using 66265-1-Ig

  U-251 cells were subjected to SDS PAGE followed by western blot with 66265-1-Ig (GLS Antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66265-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66265-1-Ig (GLS Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human gliomas using 66265-1-Ig

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 66265-1-Ig (GLS antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human gliomas using 66265-1-Ig

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 66265-1-Ig (GLS antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 66265-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66265-1-Ig (GLS antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 66265-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66265-1-Ig (GLS antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of L02 using 66265-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed L02 cells using KGA/GAC antibody (66265-1-Ig, Clone: 3A12A1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of mouse brain using 66265-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 66265-1-Ig (GLS antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HepG2 using 66265-1-Ig

  Immunofluorescent analysis of (-20℃ Ethanol ) fixed HepG2 cells using 66265-1-Ig(GLS antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: NIH/3T3 cells, rat liver tissue, fetal human brain tissue, U-251 cells, HEK-293 cells
• Positive IHC detected in: human gliomas tissue, mouse brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: mouse brain tissue, HepG2 cells, L02 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• KD/KO: See 1 publications below
• WB: See 4 publications below
• IF: See 2 publications below

Product Information

66265-1-Ig targets KGA/GAC in WB, IHC, IF, ELISA applications and shows reactivity with human, pig, mouse, rat samples.

• Tested Reactivity: human, pig, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: KGA/GAC fusion protein Ag24227
• Full Name: glutaminase
• Calculated molecular weight: 669 aa, 73 kDa
• Observed molecular weight: 58 kDa, 65 kDa
• GenBank accession number: BC038507
• Gene symbol: GLS
• Gene ID (NCBI): 2744
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

GLS, also named as GLS1 and KIAA0838, belongs to the glutaminase family. It catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine. Glutaminase-, glutamate-, and taurine-immunoreactive neurons develop neurofibrillary tangles in Alzheimer's disease(PMID:1672808).The glutaminase band in AA/C1 cells is more intense than in HT29 cells, in accordance with measurements of glutaminase activity, and had the same molecular mass of approx. 63 kDa. The bands for both cell lines are clearly different in size from both rat liver glutaminase (58 kDa) and rat kidney glutaminase (65 kDa)(PMID:12408749). It also reveals a molecular weight of 83-84 kDa as a phosphate-dependent glutaminase(PMID:447624;7512428). It has 3 isoforms named as KGA, GAM, GAC. This antibody can recognize KGA and GAC.

Publications

Species	Application	Title
		J Hepatol; OGDHL silencing promotes hepatocellular carcinoma by reprogramming glutamine metabolism.; Authors - Weiqi Dai
human	WB,IF	Mol Cell; Filamentous GLS1 promotes ROS-induced apoptosis upon glutamine deprivation via insufficient asparagine synthesis.; Authors - Bin Jiang ; KO Validated
human	WB	Int J Oncol; Cryptotanshinone suppresses tumorigenesis by inhibiting lipogenesis and promoting reactive oxygen species production in KRAS‑activated pancreatic cancer cells.; Authors - Tokio Terado
mouse	IF	Oxid Med Cell Longev; Pharmacological Inhibition of Glutaminase 1 Attenuates Alkali-Induced Corneal Neovascularization by Modulating Macrophages.; Authors - Yifan Feng
human	WB	Prostate; Prostate cancer cells demonstrate unique metabolism and substrate adaptability acutely after androgen deprivation therapy; Authors - Mikolaj J Filon
mouse	WB	J Mol Cell Cardiol; Glutamine uptake and catabolism is required for myofibroblast formation and persistence; Authors - Andrew A Gibb

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Longlong (Verified Customer) (03-01-2022)

I'm working on a glutaminase project. Great western blot results and fantastic consistency

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: human AML cell lines

KGA/GAC Antibody Western Blot validation (1:1000 dilution) in human AML cell lines (Cat no:66265-1-Ig)