- HPA019648 - Atlas Antibodies GCN1 antibody

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Enhanced validation

Submitted by: klas2
Enhanced method: Genetic validation
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Experimental details: Western blot of cell lysate from U-2 OS cells transfected with either siRNA targeting GCN1L1 or control siRNA. Lane 1: Marker (250, 130, 95, 72, 55, 36, 28, 17, 10) Lane 2: Cell lysate from U-2OS cells transfected with siRNA targeting GCN1L1 Lane 3: N/A Lane 4: Cell lysate from U-2OS cells transfected with control siRNA Right image, lane 1-4: loading control
Sample type: U-2 OS
Primary Ab dilution: 1:78
Conjugate: Horseradish Peroxidase
Secondary Ab: Secondary Ab
Secondary Ab dilution: 1:3000
Knockdown/Genetic Approaches Application: Western blot
KD Reagent Type: siRNA
Downregulation: >75%
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Prod no: s21624
KD Reagent Provider: Thermo Fisher Scientific
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 2: s21625
KD Reagent Prod no 3: s21626
KD Reagent Page 3: KD Reagent Page 3
Antibody Lot Number: R10495
Appendix: 565d676ece70c.pdf

Supportive validation

Submitted by: Atlas Antibodies (provider)
Enhanced method: Genetic validation
Main image
Experimental details: Western blot analysis in U2OS cells transfected with control siRNA, target specific siRNA probe #1 and #2, using Anti-GCN1 antibody. Remaining relative intensity is presented.

Enhanced validation

Submitted by: 55af80e3e0991
Enhanced method: Genetic validation
Main image
Experimental details: Confocal images of immunofluorescently stained human U-2 OS cells.The protein GCN1L1 is shown in green. The image to the left show cells transfected with control siRNA and the image to the right show cells where GCN1L1 has been downregulated with specific siRNA.
Sample type: U-2 OS cells
Primary Ab dilution: 1:35
Secondary Ab: Secondary Ab
Secondary Ab dilution: 1:800
Knockdown/Genetic Approaches Application: Immunocytochemistry
KD Reagent Type: siRNA
Downregulation: >50%
KD Reagent Provider: Thermo Fisher Scientific
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Prod no: s21624
KD Reagent Prod no 2: s21625
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 3: s21626
KD Reagent Page 3: KD Reagent Page 3
Antibody Lot Number: R10495
Appendix: 55b0b0933c437.pdf

Supportive validation

Submitted by: Atlas Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of human cell line U-2 OS shows localization to cytosol.
Sample type: HUMAN

Enhanced validation

Submitted by: Atlas Antibodies (provider)
Enhanced method: Orthogonal validation
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Experimental details: Immunohistochemistry analysis in human testis and liver tissues using Anti-GCN1 antibody. Corresponding GCN1 RNA-seq data are presented for the same tissues.
Sample type: HUMAN
Orthogonal method: Orthogonal validation of protein expression using IHC by comparison to RNA-seq data of corresponding target in high and low expression tissues.

Enhanced validation

Submitted by: Atlas Antibodies (provider)
Enhanced method: Independent antibody validation
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Experimental details: Immunohistochemical staining of human skeletal muscle, skin, stomach and testis using Anti-GCN1 antibody HPA019648 (A) shows similar protein distribution across tissues to independent antibody HPA018799 (B).
Antibody #2 product nr: HPA018799, HPA024367
Antibody provider: Atlas Antibodies

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human duodenum shows strong cytoplasmic positivity in glandular cells.

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human testis shows high expression.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human liver shows low expression as expected.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human cerebral cortex using Anti-GCN1 antibody HPA019648.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human colon using Anti-GCN1 antibody HPA019648.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human skin shows strong cytoplasmic positivity in squamous epithelial cells.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human skeletal muscle shows moderate cytoplasmic positivity in myocytes.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human testis shows strong cytoplasmic positivity in cells in seminiferous ducts.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
Main image
Experimental details: Immunohistochemical staining of human stomach shows strong cytoplasmic positivity in glandular cells.
Sample type: HUMAN

HPA019648