MA1-5777

antibody from Invitrogen Antibodies

Targeting: GEMIN2 SIP1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-5777

Provider

Invitrogen Antibodies

Product name

Gemin 2 Monoclonal Antibody (2E17)

Antibody type

Monoclonal

Antigen

Other

Description

MA1-5777 detects Gemin2 in human and xenopus samples.

Antibody clone number

2E17

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Gemin 2 was achieved by transfecting HeLa cells with Gemin 2 specific siRNAs (Silencer® select Product # s16118, s16120). Western blot analysis (Fig. a) was performed using whole cell extracts from the Gemin 2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Gemin 2 Monoclonal Antibody (Product # MA1-5777, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 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 Gemin 2.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HepG2 (Lane 2), MCF7 (Lane 3), PC-3 (Lane 4), K-562 (Lane 5), Jurkat (Lane 6), A549 (Lane 7), A-431 (Lane 8) and Caco-2 (Lane 9). The blot was probed with Gemin 2 Monoclonal Antibody (2E17) (Product # MA1-5777 , 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A band at ~32 kDa corresponding to Gemin 2 was observed across cell lines tested along with an additional band close to 40kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Gemin 2 was performed using 70% confluent log phase Hep G2 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 Gemin 2 Mouse Monoclonal Antibody (Product # MA1-5777) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 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 nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.