- HPA030518 - Atlas Antibodies NASP antibody

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Enhanced validation

Submitted by: klas2
Enhanced method: Genetic validation
Main image
Experimental details: Western blot of cell lysate from U-2 OS cells transfected with either siRNA targeting NASP 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 NASP 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: 1:58
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 Provider: Thermo Fisher Scientific
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Prod no: s9281
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 2: s9280
KD Reagent Page 3: KD Reagent Page 3
KD Reagent Prod no 3: s9282
Antibody Lot Number: R30241
Appendix: 566d5a59b7f1f.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-NASP antibody. Remaining relative intensity is presented.

Supportive validation

Submitted by: Atlas Antibodies (provider)
Enhanced method: Recombinant expression validation
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Experimental details: Western blot analysis in control (vector only transfected HEK293T lysate) and NASP over-expression lysate (Co-expressed with a C-terminal myc-DDK tag (~3.1 kDa) in mammalian HEK293T cells, LY400884).

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 NASP is shown in green and the microtubules in red. The image to the left show cells transfected with control siRNA and the image to the right show cells where NASP has been downregulated with specific siRNA.
Sample type: U-2 OS cells
Primary Ab dilution: 1:26
Secondary Ab: Secondary Ab
Secondary Ab dilution: 1:800
Knockdown/Genetic Approaches Application: Immunocytochemistry
KD Reagent Type: siRNA
Downregulation: >50%
KD Reagent Prod no: s9281
KD Reagent Provider: Thermo Fisher Scientific
KD Reagent Prod page: KD Reagent Prod page
KD Reagent Page 2: KD Reagent Page 2
KD Reagent Prod no 2: s9280
KD Reagent Page 3: KD Reagent Page 3
KD Reagent Prod no 3: s9282
Antibody Lot Number: R30241
Appendix: 55b5e7ed305bf.pdf

Supportive validation

Submitted by: Atlas Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of human cell line U-2 OS shows localization to nucleoplasm.
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 pancreas tissues using Anti-NASP antibody. Corresponding NASP 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
Main image
Experimental details: Immunohistochemical staining of human liver, lymph node, pancreas and testis using Anti-NASP antibody HPA030518 (A) shows similar protein distribution across tissues to independent antibody HPA028136 (B).
Antibody #2 product nr: HPA030520, HPA028136
Antibody provider: Atlas Antibodies

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Atlas Antibodies (provider)
Main image
Experimental details: Immunohistochemical staining of human lymph node using Anti-NASP antibody HPA030518.
Sample type: HUMAN

Supportive validation

Submitted by: Atlas Antibodies (provider)
Main image
Experimental details: Immunohistochemical staining of human liver using Anti-NASP antibody HPA030518.
Sample type: HUMAN

HPA030518