KGA/GAC Monoclonal antibody

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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.

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  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.

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  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.

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  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).

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  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).

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  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).

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  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).

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  IF Staining of HeLa using 66265-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using KGA/GAC antibody (66265-1-Ig, Clone: 3A12A1 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1), CL594-Phalloidin (red).

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  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).

• Featured Product
• KD/KO Validated

KGA/GAC Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, ELISA

Cat No. 66265-1-Ig

Clone No.3A12A1

Publications(11)

Reviews (2)

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, IF/ICC, IF-P, ELISA

Synonyms

GLS, KGA, GAM, GAC, EC:3.5.1.2

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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-P detected in: mouse brain tissue
• Positive IF/ICC detected in: HeLa cells, HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 6 publications below
• IHC: See 1 publications below
• IF: See 4 publications below

Product Information

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

• Tested Reactivity: human, mouse, rat, pig
• 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
• RRID: AB_2881651
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: O94925
• 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
		Nature; Cellular ATP demand creates metabolically distinct subpopulations of mitochondria; Authors - Keun Woo Ryu
		J Hepatol; OGDHL silencing promotes hepatocellular carcinoma by reprogramming glutamine metabolism.; Authors - Weiqi Dai
human	IHC,IF	Cancer Commun (Lond); Glutamine metabolic microenvironment drives M2 macrophage polarization to mediate trastuzumab resistance in HER2-positive gastric cancer; Authors - Xingbin Hu
human	WB,IF	Mol Cell; Filamentous GLS1 promotes ROS-induced apoptosis upon glutamine deprivation via insufficient asparagine synthesis.; Authors - Bin Jiang ; KO Validated
mouse	WB	Cell Rep; Glutaminase 1 deficiency confined in forebrain neurons causes autism spectrum disorder-like behaviors; Authors - Chenhui Ji
mouse	IF	Oxid Med Cell Longev; Pharmacological Inhibition of Glutaminase 1 Attenuates Alkali-Induced Corneal Neovascularization by Modulating Macrophages.; Authors - Yifan Feng

Reviews

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

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FH

Sarah-Eve (Verified Customer) (06-02-2023)

Works well

• Applications: Western Blot

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)