MA5-38467

antibody from Invitrogen Antibodies

Targeting: IL6ST CD130, GP130, sGP130

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-38467

Provider

Invitrogen Antibodies

Product name

GP130 Monoclonal Antibody (8D4D2)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

This antibody has been tested in indirect ELISA.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

8D4D2

Vial size

100 µg

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 GP130 Monoclonal Antibody (8D4D2) (Product # MA5-38467) and a band at 100 to 150 kDa corresponding to IL6ST was observed in BT-549 which is a high expression model and no band was observed in NIH:OVCAR-3 which is a low expression model . Whole cell extracts (30 µg lysate) of NIH:OVCAR-3 (Lane 1), BT-549 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis using GP130 Monoclonal Antibody (Product # MA5-38467) against human CD130 (AA: extra 73-231) recombinant protein. (Expected MW is 43.8 kDa)

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of GP130 using GP130 Monoclonal Antibody (Product # MA5-38467) in HEK293 (1) and CD130 (AA: extra 73-231)-human IgG-Fc transfected HEK293 (2) cell lysate.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis of GP130 in HeLa cells using GP130 Monoclonal Antibody (Product # MA5-38467) (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor- 555 phalloidin. Secondary antibody from Fisher (Product # 35503)

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IL6ST was performed using 70% confluent log phase BT-549 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 GP130 Monoclonal Antibody (8D4D2) (Product # MA5-38467) at 1:200 dilution 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 Golgi network and cell membrane localization. Panel e represents the low expression model NIH:OVCAR3 with no signal . Panel f 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

Immunohistochemistry (Paraffin) analysis of GP130 in paraffin-embedded cervical cancer tissues using GP130 Monoclonal Antibody (Product # MA5-38467) and DAB staining.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry analysis of GP130 in HL-60 cells using GP130 Monoclonal Antibody (Product # MA5-38467) (green) and negative control (red).