MA5-29637

antibody from Invitrogen Antibodies

Targeting: USP7 HAUSP

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-29637

Provider

Invitrogen Antibodies

Product name

USP7 Monoclonal Antibody (03)

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%). This antibody has specificity for Human USP7/HAUSP.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

3

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.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using USP7 Monoclonal Antibody (3) (Product # MA5-29637) and a ~150 kDa band corresponding to USP7 was observed across cell lines and tissues tested. Nuclear enriched extracts (30 µg lysate) of HT-29 (Lane 1), SW480 (Lane 2), A549 (Lane 3), Mouse Spleen (Lane 4), Rat Spleen (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB33001) by iBlot™ 3 Western Blot Transfer Device (Product # IB31001). The blot was probed with the primary antibody (1:1,000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580). A non specific band ~55 kDa was observed in all human cell lines. Band at 25 kDa corresponding to tissue IgG was observed in mouse spleen.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence staining of Human USP7 in A431 cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with USP7 Monoclonal Antibody (3) (Product # MA5-29637, 1:60) at 4°C overnight. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse 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 USP7 was performed using 70% confluent log phase A549 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 USP7 Monoclonal Antibody (3) (Product # MA5-29637, 1:100) in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32766), (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.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Human USP7 expression on Jurkat cells. The cells were treated according to manufacturer’s manual, stained with USP7 Monoclonal Antibody (3) (Product # MA5-29637), 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.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Human USP7 expression on Jurkat cells. The cells were treated according to manufacturer’s manual, stained with USP7 Monoclonal Antibody (3) (Product # MA5-29637), 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.