PA1-16548

antibody from Invitrogen Antibodies

Targeting: FANCD2 FA-D2, FACD, FAD, FANCD

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Chromatin Immunoprecipitation (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-16548

Provider

Invitrogen Antibodies

Product name

FANCD2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

In Western blot, this antibody should recognize a band at ~166 kDa (post-translationally modified form). Additional bands may be seen at lower molecular weights. In immunofluorescence, this has been tested in human MMC and IR-treated MEF cells.Suggested positive control: Hela whole cell extract.

Reactivity

Human, Mouse, Rat, Canine, Chicken/Avian, Zebrafish

Host

Rabbit

Isotype

IgG

Vial size

50 µL

Concentration

1 mg/mL

Storage

-20°C, Avoid Freeze/Thaw Cycles

References

Submitted references

Gene expression profiling reveals activation of the FA/BRCA pathway in advanced squamous cervical cancer with intrinsic resistance and therapy failure.

Balacescu O, Balacescu L, Tudoran O, Todor N, Rus M, Buiga R, Susman S, Fetica B, Pop L, Maja L, Visan S, Ordeanu C, Berindan-Neagoe I, Nagy V
BMC cancer 2014 Apr 8;14:246

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of FANCD2 in SiHa cells after cell exposure to IR. Samples were incubated in FANCD2 polyclonal antibody (Product # PA1-16548). Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 hours. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of FANCD2 was performed using 70% confluent log phase HeLa cells treated with 2mM of Hydroxyurea for 24 hours. 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 FANCD2 Rabbit Polyclonal Antibody (Product # PA1-16548) 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 nuclear localization. Panel e shows untreated cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of FANCD2 was performed using 70% confluent log phase HeLa cells treated with 2mM of Hydroxyurea for 24 hours. 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 FANCD2 Rabbit Polyclonal Antibody (Product # PA1-16548) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) 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 nuclear localization. Panel e shows untreated cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of FANCD2 in SiHa cells after cell exposure to IR. Samples were incubated in FANCD2 polyclonal antibody (Product # PA1-16548). Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 hours. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of FANCD2 in human prostate, glandular epithelium. Samples were incubated in FANCD2 polyclonal antibody (Product # PA1-16548). Image using the Biotin format of this antibody.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Validation of FANCD2 (A-B), RAD51 (C-D), BRCA1 (E-F), BRCA2 (G-H) and BRIP1 (I-J) protein expression in advanced squamous cervical tumor cells. Staining for FANCD2, RAD51, BRCA1 and BRIP1 for the NCR cervical samples indicated strongly positive protein expression compared with the CR cervical samples. The BRCA2 protein expression was comparable between the NCR and CR cervical samples; (x200 magnification).