PA5-21143

antibody from Invitrogen Antibodies

Targeting: NUMB C14orf41

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-21143

Provider

Invitrogen Antibodies

Product name

NUMB Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

A suggested positive control is mouse lung tissue lysate.

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of NUMB in mouse lung tissue lysate with NUMB Polyclonal Antibody (Product # PA5-21143) at (A) 0.25 and (B) 0.5 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NUMB Polyclonal Antibody (Product # PA5-21143) on whole cell extracts (30 µg lysate) of U-87 MG (Lane1), PC-3 (Lane 2), DU 145 (Lane 3), A549 (Lane 4), SH-SY5Y(Lane 5), HeLa (Lane 6), Mouse Lungs (Lane 7), Mouse Colon (Lane 8), Mouse Heart (Lane 9) and Mouse Kidney (Lane 10)and NUMB major isoform 65 kDa was observed along with other isoforms between 53-70 kDa in all samples except Mouse Colon, Heart and Kidney. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NUMB was performed using 70% confluent log phase U-87 MG 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 NUMB Polyclonal Antibody (Product # PA5-21143) at 5µg/mL 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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of NUMB in human lung tissue with NUMB Polyclonal Antibody (Product # PA5-21143) at 20 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of NUMB in human lung tissue with NUMB Polyclonal Antibody (Product # PA5-21143) at 20 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of NUMB in human lung tissue with NUMB Polyclonal Antibody (Product # PA5-21143) at 5 µg/mL.