Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling GABARAP+GABARAPL1+GABARAPL2 with Purified ab109364 at 1 : 500 dilution (0.29 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling GABARAP+GABARAPL1+GABARAPL2 with purified ab109364 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Unpurified ab109364 staining GABARAP+GABARAPL1+GABARAPL2 in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluorr^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Unpurified ab109364, at 1/500, staining GABARAP+GABARAPL1+GABARAPL2 in Human brain tissue by immunohistochemistry.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GABARAP+GABARAPL1+GABARAPL2 with purified ab109364 at 1/20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Lab

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/5000 dilution

Lane 1:

Mouse kidney lysates at 20 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 17 kDa

false

• WB

Lab

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/5000 dilution

All lanes:

Rat kidney lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 17 kDa

false

• WB

Unknown

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

All lanes:

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/1000 dilution

Lane 1:

Fetal kidney lysate at 10 µg

Lane 2:

SH-SY5Y cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

Fetal brain lysate at 10 µg

Predicted band size: 14 kDa

false

• WB

CiteAb

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Western Blotting using Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805], ab109364. Publication image from Yang, D. et al., 2022, Adv Sci (Weinh), 34783461. Legend direct from paper.

GABARAP interacts with AC3 via LIRs of AC3 in a ciliary expression‐dependent manner. A‐E) WB (A) and densitometric quantification of the expression of GABARAP (B), ATG5 (C), VPS34 (D), and Beclin1(E) in the hypothalami of AC3+/+, AC3−/−, and hAC3 mice (n = 3 mice per group). Actin served as the loading control. F) Representative IF co‐staining with AC3 and GABARAP antibodies in the VMHs of WT mice. F′) A higher magnification of the boxed region. Scale bars : F) 20 µm; F′) 5 µm. G) Representative images showing the expression levels of GABARAP and AC3 in the VMHs of VMH pIFT88‐AC3 KD mice and the controls. Scale bars : 20 µm. H) Schematic representation of GABARAP binding LIRs at aa488‐aa493 and aa958‐aa963 of AC3. I) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (LIR1 Mut), or AC3 (LIR2 Mut). LgG served as the negative control. J) Pull‐down analysis of GABARAP and AC3. K) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (296 Mut), or AC3 (465 Mut). LgG served as the negative control. L) Schematic representation of AC3 regulating GABARAP. Data represent the mean ± SEM; *p < 0.05 and **p < 0.01; one‐way ANOVA and Bonferroni pairwise comparisons.

false

• WB

CiteAb

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Western Blotting using Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805], ab109364. Publication image from Yang, D. et al., 2022, Adv Sci (Weinh), 34783461. Legend direct from paper.

GABARAP interacts with AC3 via LIRs of AC3 in a ciliary expression‐dependent manner. A‐E) WB (A) and densitometric quantification of the expression of GABARAP (B), ATG5 (C), VPS34 (D), and Beclin1(E) in the hypothalami of AC3+/+, AC3−/−, and hAC3 mice (n = 3 mice per group). Actin served as the loading control. F) Representative IF co‐staining with AC3 and GABARAP antibodies in the VMHs of WT mice. F′) A higher magnification of the boxed region. Scale bars : F) 20 µm; F′) 5 µm. G) Representative images showing the expression levels of GABARAP and AC3 in the VMHs of VMH pIFT88‐AC3 KD mice and the controls. Scale bars : 20 µm. H) Schematic representation of GABARAP binding LIRs at aa488‐aa493 and aa958‐aa963 of AC3. I) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (LIR1 Mut), or AC3 (LIR2 Mut). LgG served as the negative control. J) Pull‐down analysis of GABARAP and AC3. K) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (296 Mut), or AC3 (465 Mut). LgG served as the negative control. L) Schematic representation of AC3 regulating GABARAP. Data represent the mean ± SEM; *p < 0.05 and **p < 0.01; one‐way ANOVA and Bonferroni pairwise comparisons.

false

• WB

CiteAb

Western blot - Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (AB109364)

Western Blotting using Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805], ab109364. Publication image from Yang, D. et al., 2022, Adv Sci (Weinh), 34783461. Legend direct from paper.

GABARAP interacts with AC3 via LIRs of AC3 in a ciliary expression‐dependent manner. A‐E) WB (A) and densitometric quantification of the expression of GABARAP (B), ATG5 (C), VPS34 (D), and Beclin1(E) in the hypothalami of AC3+/+, AC3−/−, and hAC3 mice (n = 3 mice per group). Actin served as the loading control. F) Representative IF co‐staining with AC3 and GABARAP antibodies in the VMHs of WT mice. F′) A higher magnification of the boxed region. Scale bars : F) 20 µm; F′) 5 µm. G) Representative images showing the expression levels of GABARAP and AC3 in the VMHs of VMH pIFT88‐AC3 KD mice and the controls. Scale bars : 20 µm. H) Schematic representation of GABARAP binding LIRs at aa488‐aa493 and aa958‐aa963 of AC3. I) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (LIR1 Mut), or AC3 (LIR2 Mut). LgG served as the negative control. J) Pull‐down analysis of GABARAP and AC3. K) Co‐IP analysis of GABARAP and AC3 (WT), AC3 (296 Mut), or AC3 (465 Mut). LgG served as the negative control. L) Schematic representation of AC3 regulating GABARAP. Data represent the mean ± SEM; *p < 0.05 and **p < 0.01; one‐way ANOVA and Bonferroni pairwise comparisons.

false