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GLS Antibody (C-term)

Peptide Affinity Purified Rabbit Polyclonal Antibody (Pab)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue using AP8809B performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody(1:500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded Human brain tissue using AP8809B performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody(1:500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized U-251 MG cells labeling GLS with AP8809B at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-Rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing Cytoplasm staining on U-251 MG cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin(red). The nuclear counter stain is DAPI (blue).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling GLS with AP8809b at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG (1583138) secondary antibody at 1/200 dilution (green). Immunofluorescence image showing mitochondrion staining on HepG2 cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin (OI17558410) at 1/100 dilution (red). The nuclear counter stain is DAPI (blue).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling GLS with AP8809b at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG (1583138) secondary antibody at 1/200 dilution (green). Immunofluorescence image showing mitochondrion staining on HepG2 cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin (OI17558410) at 1/100 dilution (red). The nuclear counter stain is DAPI (blue).
Overlay histogram showing U-2 OS cells stained with AP8809b(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP8809b, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(OH191631) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
All lanes : Anti-GLS Antibody (C-term) at 1:2000 dilution Lane 1: human brain lysate Lane 2: mouse brain lysate Lane 3: 293T/17 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 73 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes : Anti-GLS Antibody (C-term) at 1:2000 dilution Lane 1: human brain lysate Lane 2: human kidney lysate Lane 3: mouse brain lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 73 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot analysis of lysates from human brain, mouse brain ad rat kidney tissue lysate (from left to right), using GLS Antibody (C-term)(Cat. #AP8809b). AP8809b was diluted at 1:1000 at each lane. A goat anti-rabbit IgG H&L(HRP) at 1:5000 dilution was used as the secondary antibody. Lysates at 35ug per lane.

产品详情
文献引用 : 3
实验流程
背景知识

Product info
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	IHC-P-Leica, IF, WB, FC, E
Primary Accession	O94925
Other Accession	P13264, D3Z7P3
Reactivity	Human, Mouse, Rat
Predicted	Rat
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit Ig

Additional info
Gene ID	2744
Other Names	Glutaminase kidney isoform, mitochondrial, GLS, K-glutaminase, L-glutamine amidohydrolase, GLS, GLS1, KIAA0838
Target/Specificity	This GLS antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 516-545 amino acids from the C-terminal region of human GLS.
Dilution	IHC-P-Leica~~1:500 IF~~1:25 FC~~1:25 WB~~1:1000
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	GLS Antibody (C-term) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	GLS
Synonyms	GLS1, KIAA0838
Function	Catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine. Plays a role in maintaining acid-base homeostasis. Regulates the levels of the neurotransmitter glutamate in the brain. Isoform 2 lacks catalytic activity.
Cellular Location	Isoform 1: Cytoplasm, cytosol.
Tissue Location	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

Research Areas

Heat Shock Factor 1 Epigenetically Stimulates Glutaminase-1-Dependent mTOR Activation to Promote Colorectal Carcinogenesis.
Author : Li J1,Song P1,Jiang T2,Dai D1,Wang H1,Sun J2,Zhu L2,Xu W2,Feng L2,Shin VY3,Morrison H4,Wang X5,Jin H6. Mol Ther. 2018 Jul 5;26(7):1828-1839. doi: 10.1016/j.ymthe.2018.04.014. Epub 2018 Apr 14.	29730197

CXXC4 activates apoptosis through up-regulating GDF15 in gastric cancer.
Author : Han M1,Dai D1,Yousafzai NA1,Wang F2,Wang H1,Zhou Q1,Lu H1,Xu W2,Feng L2,Jin H2,Wang X1. Oncotarget. 2017 Oct 6;8(61):103557-103567. doi: 10.18632/oncotarget.21581. eCollection 2017 Nov 28.	29262584

The oncogenic transcription factor c-Jun regulates glutaminase expression and sensitizes cells to glutaminase-targeted therapy.
Author : Lukey MJ¹, Greene KS¹, Erickson JW², Wilson KF¹, Cerione RA^1,². Nat Commun. 2016 Apr 18;7:11321. doi: 10.1038/ncomms11321.	27089238

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Sahai (1983) demonstrated phosphate-activated glutaminase (EC 3.5.1.2) in human platelets. It is the major enzyme yielding glutamate from glutamine. Significance of the enzyme derives from its possible implication in behavior disturbances in which glutamate acts as a neurotransmitter(Prusiner, 1981). High heritability of platelet glutaminase was indicated by studies of Sahai and Vogel (1983) [PubMed 6682827] who found an intraclass correlation coefficient of 0.96 for monozygotic twins and 0.53 for dizygotic twins.

REFERENCES

Swierczynski,J., et.al., Biochim. Biophys. Acta 1157 (1), 55-62 (1993)

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¥ 1,100.00

Cat# AP8809B

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GLS Antibody (C-term)

Peptide Affinity Purified Rabbit Polyclonal Antibody (Pab)

BACKGROUND

REFERENCES

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