- HPA018295 - Atlas Antibodies ATXN2 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 ATXN2 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 ATXN2 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: 1:275
Conjugate: Horseradish Peroxidase
Secondary Ab: Secondary Ab
Secondary Ab dilution: 1:3000
Knockdown/Genetic Approaches Application: Western blot
KD Reagent Type: siRNA
Downregulation: >50%
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Prod no: s12493
KD Reagent Provider: Thermo Fisher Scientific
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 2: s12492
KD Reagent Page 3: KD Reagent Page 3
KD Reagent Prod no 3: s12491
Antibody Lot Number: R07712
Appendix: 56b1cd5ad1717.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, using Anti-ATXN2 antibody. Remaining relative intensity is presented. Loading control: Anti-GAPDH.

Supportive validation

Supportive validation

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 ATXN2 is shown in green and the nucleus in blue. The image to the left show cells transfected with control siRNA and the image to the right show cells where ATXN2 has been downregulated with specific siRNA.
Sample type: U-2 OS cells
Primary Ab dilution: 1:123
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 no: s12493
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 2: s12492
KD Reagent Prod no 3: s12491
KD Reagent Page 3: KD Reagent Page 3
Antibody Lot Number: R07712
Appendix: 55b8a1efa30ee.pdf

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunofluorescent staining of human cell line U-251 MG 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 skeletal muscle tissues using HPA018295 antibody. Corresponding ATXN2 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 cerebellum, cerebral cortex, skeletal muscle and testis using Anti-ATXN2 antibody HPA018295 (A) shows similar protein distribution across tissues to independent antibody HPA020339 (B).
Antibody #2 product nr: HPA020339
Antibody provider: Atlas Antibodies

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human cerebral cortex shows strong cytoplasmic positivity in neuronal 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 skeletal muscle 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 colon using Anti-ATXN2 antibody HPA018295.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human kidney using Anti-ATXN2 antibody HPA018295.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human skeletal muscle shows no positivity in myocytes as expected.
Sample type: HUMAN

Supportive validation

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

Supportive validation

Submitted by: Atlas Antibodies (provider)
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Experimental details: Immunohistochemical staining of human cerebral cortex shows moderate to strong cytoplasmic positivity in neurons.
Sample type: HUMAN

Supportive validation

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

HPA018295