PA5-29151

antibody from Invitrogen Antibodies

Targeting: TIGAR C12orf5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

PA5-29151

Provider

Invitrogen Antibodies

Product name

TIGAR Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: 293T, A431, H1299, HeLaS3, HepG2, Molt-4, Raji. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

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.

References

Submitted references

TIGAR inclusion pathology is specific for Lewy body diseases.

López KLR, Simpson JE, Watson LC, Mortiboys H, Hautbergue GM, Bandmann O, Highley JR
Brain research 2019 Mar 1;1706:218-223

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 MCF7 (Lane 1), MCF7 treated with Doxorubicin (0.5uM for 24hr) (Lane 2), HeLa (Lane 3) and U-87 MG (Lane 4). The blot was probed with Anti-TIGAR Polyclonal Antibody (Product # PA5-29151, 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 30 kDa band corresponding to TIGAR was observed across cell lines tested and was enhanced upon Doxorubicin treatment in MCF7 cell line.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using TIGAR Polyclonal Antibody (Product # PA5-29151). Sample (30 µg whole cell lysate). A: Hep G2. B: MOLT4. 12% SDS PAGE. TIGAR Polyclonal Antibody (Product # PA5-29151) diluted at 1:3,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of TIGAR was performed using 70% confluent log phase Hep G2 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 TIGAR Polyclonal Antibody (Product # PA5-29151) at 1:100 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) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). 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.