PA5-22048

antibody from Invitrogen Antibodies

Targeting: RAG2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-22048

Provider

Invitrogen Antibodies

Product name

RAG2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: Molt-4, rat thymus.

Concentration

1.01 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of Jurkat (Lane 1), Ramos (Lane 2), Hep G2 (Lane 3), HeLa (Lane 4) and MOLT-4 (Lane 5). The blot was probed with Anti-RAG2 Polyclonal Antibody (Product # PA5-22048, 1:500 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 55 kDa band corresponding to RAG2 was observed in all cell lines along with additional uncharacterized bands (*) between ~70 kDa and ~110 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using RAG2 Polyclonal Antibody (Product # PA5-22048). Sample (30 µg of whole cell lysate). Lane A: Molt-4 . 7.5% SDS PAGE. RAG2 Polyclonal Antibody (Product # PA5-22048) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

RAG2 Polyclonal Antibody detects RAG2 protein by Western blot analysis. A. 50 µg Rat thymus lysate/extract.7.5 % SDS-PAGE. RAG2 Polyclonal Antibody (Product # PA5-22048) dilution: 1:1,000.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

RAG2 Polyclonal Antibody detects RAG2 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: RAG2 protein stained by RAG2 Polyclonal Antibody (Product # PA5-22048) diluted at 1:500. Red: Phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar = 10 µm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of RAG2 was performed using 70% confluent log phase HeLa 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 RAG2 Polyclonal Antibody (Product # PA5-22048) at 1:200 dilution 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) 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 predominant nuclear localization along with weak cytoplasmic signal. 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 DLD1 Xenograft, using RAG2 (Product # PA5-22048) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.