MA5-29587

antibody from Invitrogen Antibodies

Targeting: STUB1 CHIP, HSPABP2, NY-CO-7, SDCCAG7, UBOX1

Western blot (Data presented on provider website)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-29587

Provider

Invitrogen Antibodies

Product name

STUB1 Recombinant Rabbit Monoclonal Antibody (034)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

This product is preservative free. It is recommended to add sodium azide to avoid contamination (final concentration 0.05%-0.1%). Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire. This antibody has specificity for Human STUB1.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

34

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence staining of STUB1 in MCF7 cells. Cells were fixed with 4% PFA, permeabilzed with 0.1% Triton X-100 in PBS, blocked with 10% serum, and incubated with STUB1 Recombinant Rabbit Monoclonal Antibody (34) (Product # MA5-29587, 1:300) at 4°C overnight. Then cells were stained with the Alexa Fluor®488-conjugated Goat Anti-rabbit IgG secondary antibody (green). Positive staining was localized to cytoplasm and nucleus.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of STUB1 was performed using 70% confluent log phase MCF7 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 STUB1 Recombinant Rabbit Monoclonal Antibody (Product # MA5-29587) at 1:100 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). Panel d represents the merged image showing Nucleus and Cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Human STUB1 expression on Hela cells. The cells were treated according to manufacturer’s manual, stained with STUB1 Recombinant Rabbit Monoclonal Antibody (34) (Product # MA5-29587), then a FITC-conjugated Secondary antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

STUB1 Immunoprecipitation using: Lane A: 0.5 mg Hela Whole Cell Lysate, Lane B: 0.5 mg 293T Whole Cell Lysate, Lane C: 0.5 mg MCF-7 Whole Cell Lysate 2 µL with STUB1 Recombinant Rabbit Monoclonal Antibody (34) (Product # MA5-29587) and 15 µL of 50 % Protein G agarose. Primary antibody: STUB1 Recombinant Rabbit Monoclonal Antibody (34), at 1:500 dilution. Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 35 kDa. Observed band size: 35 kDa.