- PA5-28991 - Invitrogen Antibodies MAD1L1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Mad 1 using Various whole cell extracts (30 µg). Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Mad 1 polyclonal antibody (Product # PA5-28991) at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of MAD1 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a MAD1 Polyclonal Antibody (Product # PA5-28991) at a dilution of 1:5,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using MAD1 Polyclonal Antibody (Product # PA5-28991). Sample (30 µg of whole cell lysate). A: Molt-4. 7.5% SDS PAGE. MAD1 Polyclonal Antibody (Product # PA5-28991) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MAD1 Polyclonal Antibody detects MAD1L1 protein by western blot analysis. A. 30 µg Neuro2A whole cell lysate/extract. B. 30 µg GL261 whole cell lysate/extract. C. 30 µg C8D30 whole cell lysate/extract. D. 30 µg NIH-3T3 whole cell lysate/extract. E. 30 µg BCL-1 whole cell lysate/extract. F. 30 µg Raw264.7 whole cell lysate/extract.7.5% SDS-PAGE. MAD1 Polyclonal Antibody (Product # PA5-28991) dilution: 1:3,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MAD1 Polyclonal Antibody detects MAD1L1 protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract. B. 30 µg Rat2 whole cell lysate/extract.7.5% SDS-PAGE. MAD1 Polyclonal Antibody (Product # PA5-28991) dilution: 1:3,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of MAD1 was achieved by transfecting HCT116 cells with MAD1 specific siRNAs (Silencer® select Product # s228313, s15906). Western blot analysis (Fig. a) was performed using whole cell extracts (1% SDS) from the MAD1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with MAD1 Polyclonal Antibody (Product # PA5-28991, 1:4000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to MAD1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of HCT 116 (Lane 1), MCF-7 (Lane 2), MDA-MB-231 (Lane 3), SK-BR-3 (Lane 4), KARPAS 299 (Lane 5) and HT-29 (Lane 6). The blot was probed with MAD1 Polyclonal Antibody (Product # PA5-28991, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 83kDa band corresponding to MAD1 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of MAD1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: MAD1 Polyclonal Antibody (Product # PA5-28991) diluted at 1:500. Red: HEC1, a kinetochore marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of MAD1 was performed using 70% confluent log phaseMCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MAD1 Polyclonal Antibody (Product # PA5-28991) at 5 µg/mL concentration in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded SW480 Xenograft, using Mad 1 (Product # PA5-28991) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of MAD1 was performed in HeLa whole cell extracts using 5 µg of MAD1 Polyclonal Antibody (Product # PA5-28991). Samples were transferred to a membrane and probed with MAD1 Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-28991