KGA/GAC Polyclonal antibody

• 

  WB analysis of A549 using 12855-1-AP

  WB result of KGA/GAC antibody (12855-1-AP; 1:8000; incubated at room temperature for 1.5 hours) with sh-Control and sh-KGA/GAC transfected A549 cells.

• 

  WB analysis of HEK-293 using 12855-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody at dilution of 1:600 incubated at 4 degree celsius over night.

• 

  WB analysis using 12855-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:30000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 12855-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:12000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 12855-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 12855-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse kidney using 12855-1-AP

  mouse kidney tissue were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 12855-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 12855-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody at dilution of 1:600 incubated at room temperature for 1.5 hours.

• 

  WB analysis of COLO 320 using 12855-1-AP

  COLO 320 cells were subjected to SDS PAGE followed by western blot with 12855-1-AP (KGA/GAC antibody at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  IP experiment of mouse brain using 12855-1-AP

  IP result of anti-KGA/GAC (IP:12855-1-AP, 4ug; Detection:12855-1-AP 1:5000) with mouse brain tissue lysate 2400 ug.

• 

  IP experiment of mouse brain using 12855-1-AP

  IP result of anti-KGA/GAC (IP:12855-1-AP, 4ug; Detection:12855-1-AP 1:5000) with mouse brain tissue lysate 2400 ug.

• 

  IHC staining of human gliomas using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 12855-1-AP (KGA/GAC 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 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 12855-1-AP (KGA/GAC antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human gliomas using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 12855-1-AP (KGA/GAC 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 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 12855-1-AP (KGA/GAC antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human kidney using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 12855-1-AP (KGA/GAC antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human kidney using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 12855-1-AP (KGA/GAC antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human brain using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain using 12855-1-AP (KGA/GAC antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human brain using 12855-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain using 12855-1-AP (KGA/GAC antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of L02 using 12855-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed L02 cells using KGA/GAC antibody (12855-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

• 

  IF Staining of L02 using 12855-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed L02 cells using KGA/GAC antibody (12855-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 12855-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human KGA/GAC (12855-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP, Clone: ) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

KGA/GAC Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 12855-1-AP

Publications(110)

Reviews (1)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (2)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA

Synonyms

GLS, KGA, AAD20, EC:3.5.1.2, GAC

---

Tested Applications

• Positive WB detected in: mouse brain tissue, A549 cells, COLO 320 cells, HEK-293 cells, mouse kidney tissue, rat brain tissue, rat liver tissue
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human gliomas tissue, human kidney tissue, human 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/ICC detected in: L02 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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 11 publications below
• WB: See 97 publications below
• IHC: See 7 publications below
• IF: See 19 publications below
• IP: See 4 publications below
• CoIP: See 1 publications below

Product Information

12855-1-AP targets KGA/GAC in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, bovine, cat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: KGA/GAC fusion protein Ag3505
• 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_2110381
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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
mouse,human	WB,IP	Cell Res; AMPK targets PDZD8 to trigger carbon source shift from glucose to glutamine; Authors - Mengqi Li
human	WB	Nature; CFIm25 links alternative polyadenylation to glioblastoma tumour suppression.; Authors - Chioniso P Masamha
mouse	IF	Cell; Glia-to-Neuron Conversion by CRISPR-CasRx Alleviates Symptoms of Neurological Disease in Mice.; Authors - Haibo Zhou
mouse	WB	Cell; Systemic elevation of PTEN induces a tumor-suppressive metabolic state.; Authors - Garcia-Cao Isabel I
	WB	Cell Metab; Rewiring of Glutamine Metabolism Is a Bioenergetic Adaptation of Human Cells with Mitochondrial DNA Mutations.; Authors - Qiuying Chen
mouse	WB	Nat Metab; Quantitative analysis of metabolic fluxes in brown fat and skeletal muscle during thermogenesis; Authors - Grace Park

Reviews

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

---

FH

Kishor (Verified Customer) (01-13-2020)

Anti body works for human and mouse proteins. 1:1000 dilution in BSA+TBST is good.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: H69, HUCCT-1 and Rat Liver tissue

KGA/GAC Antibody Western Blot, validation (1:1000 dilution) in H69, HUCCT-1 and Rat Liver tissue (Cat no:12855-1-AP)