KGA-Specific Monoclonal antibody

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  WB analysis of mouse brain using 66265-2-Ig

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 66265-2-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66265-2-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66265-2-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of COLO 320 using 66265-2-Ig

  COLO 320 cells were subjected to SDS PAGE followed by western blot with 66265-2-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HSC-T6 using 66265-2-Ig

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 66265-2-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of NIH/3T3 using 66265-2-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 66265-2-Ig (GLS antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse brain using 66265-2-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66265-2-Ig (GLS antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 66265-2-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66265-2-Ig (GLS antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse brain using 66265-2-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using KGA-Specific antibody (66265-2-Ig, Clone: 1B11A11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of mouse brain using 66265-2-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using KGA-Specific antibody (66265-2-Ig, Clone: 1B11A11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of A549 using 66265-2-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed A549 cells using KGA-Specific antibody (66265-2-Ig, Clone: 1B11A11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1).

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

Cat No. 66265-2-Ig

Clone No.1B11A11

Publications(1)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

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: mouse brain tissue, HEK-293 cells, COLO 320 cells, HSC-T6 cells, NIH/3T3 cells
• Positive IHC detected in: 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: A549 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)-P: IF-P : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

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

• WB: See 1 publications below
• IF: See 1 publications below

Product Information

66265-2-Ig targets KGA-Specific in WB, IHC, IF/ICC, IF-P, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag24227

  Product name: Recombinant human GLS protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 370-669 aa of BC038507

  Sequence: YVGFSNATFQSERESGDRNFAIGYYLKEKKCFPEGTDMVGILDFYFQLCSIEVTCESASVMAATLANGGFCPITGERVLSPEAVRNTLSLMHSCGMYDFSGQFAFHVGLPAKSGVAGGILLVVPNVMGMMCWSPPLDKMGNSVKGIHFCHDLVSLCNFHNYDNLRHFAKKLDPRREGGDQRVKSVINLLFAAYTGDVSALRRFALSAMDMEQRDYDSRTALHVAAAEGHVEVVKFLLEACKVNPFPKDRWNNTPMDEALHFGHHDVFKILQEYQVQYTPQGDSDNGKENQTVHKNLDGLL

• Full Name: glutaminase
• Calculated Molecular Weight: 669 aa, 73 kDa
• Observed Molecular Weight: 65 kDa
• GenBank Accession Number: BC038507
• Gene Symbol: GLS
• Gene ID (NCBI): 2744
• RRID: AB_2881652
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G 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.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 produced by alternative splicing named as KGA, GAM, GAC. This antibody is specific to KGA.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB,IF	Cell Death Discov; O-GlcNAcylation of glutaminase isoform KGA inhibits ferroptosis through activation of glutaminolysis in hepatoblastoma; Authors - Sijia Fang