PA5-31111

antibody from Invitrogen Antibodies

Targeting: CD226 DNAM-1, DNAM1, PTA1, TLiSA1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-31111

Provider

Invitrogen Antibodies

Product name

CD226 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, Jurkat, Raji. Predicted reactivity: Rhesus Monkey (92%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Rat

Host

Rabbit

Isotype

IgG

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 analysis was performed on whole cell extracts (30 µg lysate) of Platelets (Lane 1), PC-12 (Lane 2), HeLa (Lane 3) and HEK-293 (Lane 4). The blot was probed with Anti-CD226 Polyclonal Antibody (Product # PA5-31111, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 60 kDa band corresponding to CD226 was detected in Platelets and PC-12 and lower level of expression was observed in HeLa and HEK-293 which are reported negative for CD226 expression.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using CD226 Polyclonal Antibody (Product # PA5-31111). Sample (30 µg of whole cell lysate). Lane A: Jurkat. 10% SDS PAGE. CD226 Polyclonal Antibody (Product # PA5-31111) diluted at 1:2,000.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CD226 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD226 Polyclonal Antibody (Product # PA5-31111) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ 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, 1:300). Panel d represents the merged image showing membranous 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

Immunohistochemical analysis of paraffin-embedded CL1-5 xenograft, using CD226 (Product # PA5-31111) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.