MA5-34787

antibody from Invitrogen Antibodies

Targeting: TYROBP DAP12, KARAP, PLO-SL, PLOSL

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-34787

Provider

Invitrogen Antibodies

Product name

TYROBP Recombinant Rabbit Monoclonal Antibody (JG38-70)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Positive Control: THP-1, K562, human lung cancer tissue.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

JG38-70

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles, store in dark

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of DAP12 in THP-1 cell. Samples were incubated with DAP12 monoclonal antibody (Product # MA5-34787), at a dilution of 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using TYROBP Recombinant Rabbit Monoclonal Antibody (JG38-70) (Product # MA5-34787) and a ~12 kDa band corresponding to Tyrobp was observed in THP1 and Higher expression was observed in THP1 differentiated to Macrophages except MCF7, HACAT and RAW 264.7 were reported to be negative. Whole cell Lysate (30 µg lysate) of THP-1 (Lane 1), U-937 (Lane 1), THP-1 (Lane 2), THP-1 (Lane 3), MCF7 (Lane 3), MCF7 (Lane 4), HaCaT (Lane 4), HaCaT (Lane 5), RAW 264.7 (Lane 5), RAW 264.7 (Lane 6) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX), 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 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).A single non specific band was observed (*).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of TYROBP is performed using THP-1 cells and THP1 differentiated to Macrophages. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TYROBP Recombinant Rabbit Monoclonal Antibody (JG38-70) (Product # MA5-34787) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32790, 1:2000 dilution), 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 withAlexa Fluor™ 488 Phalloidin (Product # A12379, 1:300). Panel d represents the merged image showing Plasma Membrane localization. Panel e represents THP1 undifferentiated cells. 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 analysis of DAP12 in paraffin-embedded human lung cancer tissue. Samples were incubated with DAP12 monoclonal antibody (Product # MA5-34787), and followed by hematoxylin.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of DAP12 in K562 cells (purple) compared with an unlabelled control (cells without incubation with primary antibody; yellow). Samples were incubated with DAP12 monoclonal antibody (Product # MA5-34787) at a dilution of 1:100, followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG.