Anti-APG5L/ATG5 抗体 [EPR1755(2)] - Mouse IgG1 (Chimeric)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Immunohistochemical analysis of paraffin-embedded (A) Parental HAP1 (h tissue labeling APG5L/ATG5 with ab307842 at 1/2000 (0.415 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on (A) parental HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet, no staining on (B) APG5L/ATG5 knockout HAP1 (HO010083) cell pellet.The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on human ovarian cancer.The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on mouse colon.The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on rat colon.The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBSFalse colour image of Western blot : Anti-APG5L/ATG5 antibody [EEPR1755(2)] (ab307842) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab307842 was shown to bind specifically to APG5L/ATG5 . A band was observed at 55 kDa in wild-type HAP1 cell lysates with no signal observed at this size in APG5L/ATG5 knockout cell line. To generate this image, wild-type and APG5L/ATG5 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. Exposure time :

All lanes:

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

APG5L/ATG5 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte)whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast)whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Observed band size: 55 kDa

false

• WB

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Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (AB307842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lanes 1-3 : 48 seconds；Lanes 4-9 : 92 seconds；Lanes 10-13 : 37 seconds

All lanes:

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842) at 1/1000 dilution

Lane 1:

HT-1080 (human fibrosarcoma epithelial cell)whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell)whole cell lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 10 µg

Lane 4:

Mouse heart tissue lysate at 10 µg

Lane 5:

Mouse kidney tissue lysate at 10 µg

Lane 6:

Mouse spleen tissue lysate at 10 µg

Lane 7:

Rat brain tissue lysate at 10 µg

Lane 8:

Rat heart tissue lysate at 10 µg

Lane 9:

C6 (rat glial tumor glial cell)whole cell lysate at 10 µg

Lane 10:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage)whole cell lysate at 10 µg

Lane 11:

PC-12 (rat adrenal gland pheochromocytoma)whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/20000 dilution

Observed band size: 55 kDa

false

Exposure time: 48s