Anti-GFP antibody

• IHC-FrFl

AbReview19496****

Immunohistochemistry - Free Floating - Anti-GFP antibody (AB290)

GFP Immunohistochemistry (Free Floating) analysis of mouse brain tissue sections with ab290. Tissue was fixed with 4% PFA frozen 30 μm sections were blocked for 1 hour at room temperature with 10% normal goat serum + donkey anti-mouse IgG Fab fragments (0.1 mg/ml). Sections were incubated with the primary antibody at a dilution of 1/1000 in TBS + 0.25% Triton-X for 16 hours at 4°C. A Cy2^®-conjugated donkey anti-rabbit IgG (H+L) at a dilution of 1/200 was used as the secondary antibody.

Image shows anti-NeuN (red) DAPI (blue) and anti-GFP staining of GFP-cre (green yellow with NeuN colocalization).

This image is courtesy of an Abreview submitted by Judith Kranz

• IP

AbReview16535****

Immunoprecipitation - Anti-GFP antibody (AB290)

GFP immunoprecipitation with GFP antibody ab290 in human HEK293 cells transfected with Annexin1-GFP. 25μg of cell lysate was incubated with the primary antibody and matrix (Protein G) in 1% TX-100, 10% glycerol, 1X PBS for 16 hours at 4°C. For Western blotting anti-rabbit HRP conjugated secondary antibody was used at a dilution at 1/5000.

Lane 1 : Lysate of HEK293 cells expressing Annexin1-GFP fusion protein.
Lane 2 : IP with anti-GFP.
Lane 3 : Not bound fraction.

All lanes:

Immunoprecipitation - Anti-GFP antibody (ab290)

Predicted band size: 27 kDa

false

This image is courtesy of an Abreview submitted by Vladimir Milenkovic

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (AB290)

GFP staining with GFP antibody ab290 in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab290 at 1/200 dilution overnight at +4°C followed by incubation with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) for 1 hour at 1μg/ml.

Under identical experimental conditions when compared to the basal level of GFP expression in transfected NIH3T3 cells the cells upon which ab290 was applied gave a stronger signal in the 488 channel indicating that ab290 is binding to GFP and therefore eliciting signal amplification.

ab290 was also applied to non-GFP-transfected NIH3T3 cells which produced no positive staining indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (AB290)

Bone marrow-derived infiltrating cells in the stromal tissue of gastric intraepithelial tumor traced by GFP direct fluorescence.

(A) Normal tissues of the glandular stomach of a regular GFP(−) control mouse. (B) Normal tissues of the glandular stomach of a GFP(+) transgenic control mouse; (C E D F) An induced gastric intraepithelial neoplasia (GIN) in a bone marrow transplanted mouse. GFP(+) BMDCs tracked with direct fluorescence localized in the GIN stromal tissue are shown in C and E. The same GIN lesion slide stained by H&E after the fluorescence observation are shown in D and F. DAPI (A–C and E) and hematoxylin (D and F) are used to visualize nuclei respectively. Locations of the images C and D in the images E and F and the image E in the image F are marked in the corresponding color. The gastric glands and stromal cells are also labeled.

Image from Yang C et al., PLoS One. 2013;8(11):e79615. Fig 2.; doi:10.1371/journal.pone.0079615. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• ICC

Unknown

Immunocytochemistry - Anti-GFP antibody (AB290)

GFP immunofluorescence with GFP antibody ab290, images showing similar localization of Yes-GFP (first 10 aa's of Yes PTK fused to the N-terminus of GFP) to full length Yes PTK. A : Distribution of Yes detected using mouse anti-Yes Ab followed by Texas Red-conjugated anti-mouse Ab. B : Chimeric GFP's detected using rabbit anti-GFP Ab (Abcam ab290) followed by FITC-conjugated anti-rabbit Ab.

Image kindly provided by L.G. Berthiaume. Taken from J. McCabe and L.G. Berthiaume, Functional Roles for Fatty Acylated Amino-terminal Domains in Subcellular Localization, Molecular Biology of the Cell 10 : 3771-3786, 1999

• IHC-P

AbReview9130****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (AB290)

GFP staining with ab290 in dog hearts (Adv-GFP injection) tissue sections by IHC-P. Sections were PFA fixed and subjected to heat mediated antigen retrieval in citric acid (Ph6.0 0.05% Tween20) prior to blocking with 10% serum for 30 mins at 37°C. The primary antibody was diluted 1/1000 in PBS and incubated with the sample for 1 hour at 25°C. A HRP conjugated secondary like Goat Anti-Rabbit IgG H&L (HRP) (ab205718) was used.

This image is courtesy of an anonymous Abreview

• ICC/IF

AbReview53061****

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (AB290)

GFP staining with GFP antibody ab290 in U2OS cells expressing TRF2-GFP fusion protein by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde permeabilized with NP40 and blocked with 3% BSA for 1 hour at 21°C. Samples were incubated with the primary antibody (1/1000 in PBS + 3% BSA) for 12 hours at 4°C. An Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at a dilution of 1/500 was used as the secondary antibody.

Green - GFP.
Blue - DAPI.

This image is courtesy of an anonymous Abreview

• IP

AbReview25001****

Immunoprecipitation - Anti-GFP antibody (AB290)

GFP immunoprecipitation with ab290 in HEK293 nuclear lysate expressing GFP. 20μg of lysate was incubated with primary antibody (1 μg/mg lysate) and matrix (Protein G) for 16 hours at 4°C in AFC low salt buffer. For western blotting ab290 (1/5000) was used to confirm successful immunoprecipation.

Lane 1 : HEK293 nuclear lysate expressing GFP input.
Lane 2 : IP of HEK293 nuclear lysate expressing GFP.
Lane 3 : Cells with no GFP.

All lanes:

Immunoprecipitation - Anti-GFP antibody (ab290)

Predicted band size: 27 kDa

false

This image is courtesy of an Abreview submitted by William Hung

• WB

Lab

Western blot - Anti-GFP antibody (AB290)

Secondary antibody - goat anti-rabbit HRP preadsorbed (ab97080)

All lanes:

Western blot - Anti-GFP antibody (ab290) at 1/2500 dilution

All lanes:

Western blot - Recombinant A. victoria GFP protein (<a href='/en-us/products/proteins-peptides/recombinant-a-victoria-gfp-protein-ab84191'>ab84191</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

true

Exposure time: 30s

• WB

AbReview7559****

Western blot - Anti-GFP antibody (AB290)

Blocked with 5% milk for 1 hour at 20°C.

Incubated with the primary antibody for 18 hours at 4°C in TBS containing 2% milk and 1% Tween.

Predicted MW of Eml4 ~ 120 kDa.

All lanes:

Western blot - Anti-GFP antibody (ab290) at 1/5000 dilution

Lane 1:

COS7 whole cell lysate - transfected with GFP-Eml4 at 20 µg

Lane 2:

COS7 whole cell lysate - transfected with GFP at 20 µg

Secondary

All lanes:

HRP-conjugated pig anti-rabbit IgG at 1/5000 dilution

Predicted band size: 27 kDa

Observed band size: 150 kDa,30 kDa

true

Exposure time: 10s

This image is courtesy of an Abreview submitted by S Houtman

• WB

AbReview47614****

Western blot - Anti-GFP antibody (AB290)

Blocked with 5% milk for 1 hour at 23°C.

Incubated with the primary antibody for 16 hours at 4°C.

All lanes:

Western blot - Anti-GFP antibody (ab290) at 1/5000 dilution

Lane 1:

LNCaP whole cell lysate - pEGFP empty vector at 20 µg

Lane 2:

LNCaP whole cell lysate - pEGFP-PKD1 transfected at 20 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/10000 dilution

Predicted band size: 27 kDa

true

Exposure time: 10s

This image is courtesy of an anonymous Abreview

• WB

AbReview76952****

Western blot - Anti-GFP antibody (AB290)

All lanes:

Western blot - Anti-GFP antibody (ab290) at 1/2000 dilution

All lanes:

HEK293 whole cell lysate at 5 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/10000 dilution

false

This image is courtesy of an anonymous Abreview