PA3-16829

antibody from Invitrogen Antibodies

Targeting: NUMA1

Provider product page for PA3-16829

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Immunocytochemistry [3]
Other assay [3]

Submit

Product number: PA3-16829 - Provider product page
Provider: Invitrogen Antibodies
Product name: NuMA Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Suggested positive control: Hela whole cell extract.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: Conc. Not Determined
Storage: -20°C or -80°C if preferred

NUMA1 protein structure - PA3-16829 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of NuMA in Human HeLa cells fixed with 3.5% formaldehyde. Samples were incubated in NuMA polyclonal antibody (Product # PA3-16829) using a dilution of 1:1000. The staining pattern shows the typical pattern for NuMA in the cell nucleus during interphase and on spindles during mitosis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of NuMA in Human HeLa cells fixed with 3.5% formaldehyde. Samples were incubated in NuMA polyclonal antibody (Product # PA3-16829) using a dilution of 1:1000. The staining pattern shows the typical pattern for NuMA in the cell nucleus during interphase and on spindles during mitosis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of NuMA was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with NuMA Polyclonal Antibody (Product # PA3-16829) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 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)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. WDR62-katanin regulates spindle organization and dynamics. (A) Immunofluorescence staining for alpha-tubulin and DAPI in control and indicated knockout (KO) HeLa cells. Maximum intensity projections of Z series with 30 stacks at 0.11-um steps are shown. (B and C) Quantification of astral MT number and length as shown in A. n = 28, 24, and 20 spindle poles for control, p80 knockout, and WDR62 knockout, respectively. (D) Left: Immunofluorescence staining for NuMA, gamma-tubulin, and DAPI in control and the indicated knockout HeLa cells. Insets show enlargements of the boxed areas. Right: Normalized line-scan intensity profiles of NuMA (green) and gamma-tubulin (red) corresponding to the white dashed lines in left images. (E) 3D reconstruction of images shown in D. The length of the grid in the images is 3.65 um. (F) Quantification of centrosome-to-centrosome distance and pole-to-pole distance (see Materials and methods for details) in control and indicated knockout HeLa cells. n = 40 cells for all conditions. (G) Left: Schematic representation of the distance between centrosome and pole (denoted by double-headed arrow), as measured from the center of gamma-tubulin (gamma-tub) spot (red) to the distal edge of NuMA spot (green). Right: Quantification of the distance between centrosome and pole in control and indicated knockout HeLa cells. n = 80 poles for all conditions. (H) Left: Schematic representation of outside and inside. The centers of two semicircular ROIs were

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure S2. Overview of WDR62 deletion mutants used in this study and the rescue of spindle defects by WT WDR62-GFP but not its katanin-binding deficient mutants. (A) Schematic illustration of the functional domains of WDR62 and summary of all the deletion mutants used in this study. WD40 domain (WD; light blue): MT binding and Aurora A binding; M1 (dark blue): p80 binding; M3: (green): autoinhibitory domain that interacts with WD40 domain. The horizontal lines were drawn to indicate the region spanned by the corresponding fragments (M1: dark blue; M3: green; others: black). The vertical dashed lines were drawn to indicate the boundaries of fragments M1 and M3. Note that JNK phosphorylation site T1053 (red) was located within the autoinhibitory M3 fragment. M, middle region; CC, coiled-coil. (B) Immunofluorescence staining of alpha-tubulin and DAPI in WDR62 knockout (KO) HeLa cells or the knockout cells transiently transfected with GFP-tagged WT WDR62 or its indicated mutants. Maximum intensity projections of Z series with 30 stacks at 0.11-um steps are shown. (C and D) Quantification of the number and length of astral MTs shown in B. From left to right, n = 20, 20, 22, 20, and 20 spindle poles. (E) Immunofluorescence staining of NuMA and gamma-tubulin in WDR62 knockout HeLa cells or the knockout cells transiently transfected with GFP-tagged WT WDR62 or its indicated mutants. Insets show enlargement of boxed areas. (F and G) Quantification of ""outside"" versus total intensity