重组Anti-IGF2BP1/IMP1抗体[EPR26408-18]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining ABHD2 with ab324946 at a 1/2000 dilution, ab315800 anti-CYP51A1 used at 1/2000 dilution ( 0.26 μg/ml) and ab290736 anti-IGF2BP1/IMP1 used at 1/2000 dilution ( 0.284 μg/ml).

Panel A : anti-ABHD2 (green; Opal™520), anti-CYP51A1 (magenta; Opal™690), anti-IGF2BP1/IMP1 (gray; Opal™570) on human testis.

Panel B : anti-ABHD2 staining spermatocytes in human testis.

Panel C : anti-CYP51A1 staining leydig cells in human testis.

Panel D : anti-IGF2BP1/IMP1 staining spermatogonia in mouse colon.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324946, ab315800 and ab290736 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized Caco-2 (human colorectal adenocarcinoma epithelial cell) cells lebelling IGF2BP1/IMP1 with ab290736 at 1/50 (11.34 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in Caco-2 cell line. Negative control : U-2 OS (PMID : 23069990). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling IGF2BP1/IMP1 with ab290736 at 1/2000 (0.284 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Positive staining on human testis (PMID : 28333300) . The section was incubated with ab290736 for 30 mins at room temperature.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^TM Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human testis tissue. Panel A : Merged staining of anti-IGF2BP1/IMP1 (green; Opal™690), anti-TPTE (gray; Opal™520) and anti-SAGE1 (red; Opal™570) on human testis. Panel B : Anti-IGF2BP1/IMP1 stained on cytoplasm of spermatogonia. Panel C : Anti-TPTE stained on spermatocytes. Panel D : Anti-SAGE1 stained on nucleus of spermatogonia. The section was incubated in three rounds of staining : in the order of ab290736, ab250148, and ab233388 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-2 OS (human bone osteosarcoma epithelial cell, Left) / Caco-2 (human colorectal adenocarcinoma epithelial cell, Right) cells labelling IGF2BP1/IMP1 with ab290736 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : U-2 OS (PMID : 26917013)

• mIHC

Lab

Multiplex immunohistochemistry - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human testis tissue. Panel A : Merged staining of anti-AKAP4 (magenta; Opal™690), anti-TPTE (green; Opal™520) and anti-IGF2BP1/IMP1 (red; Opal™570) on human testis. Panel B : Anti-IGF2BP1/IMP1 stained on spermatogonia. Panel C : Anti-TPTE stained on spermatocytes. Panel D : Anti-AKAP4 stained on spermatids. The section was incubated in three rounds of staining : in the order of ab238887, ab250148, and ab290736 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling IGF2BP1/IMP1 with ab290736 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling IGF2BP1/IMP1 with ab290736 at 1/2000 (0.284 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab290736 for 30 mins at room temperature.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^TM Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized NIH/3T3 (mouse embryonic fibroblast) cells lebelling IGF2BP1/IMP1 with ab290736 at 1/50 (11.34 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling IGF2BP1/IMP1 with ab290736 at 1/2000 (0.284 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab290736 for 30 mins at room temperature.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^TM Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Blocking / Diluent buffer and concentration : 5% NFDM/TBST

Exposure time :

Lane 1-3 : 10 seconds
Lane 4 : 26 seconds

Negative control : U-2 OS (PMID : 26917013)

All lanes:

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (ab290736) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 63 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Blocking / Dilution buffer and concentration : 5% NFDM/TBST

The expression profile observed is consistent with that described in the literature (PMID : 26917013).
Negative control : mouse liver, mouse kidney (PMID : 26917013).
Two unidentified bands around 37 kDa were observed.

All lanes:

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (ab290736) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 63 kDa

Observed band size: 65 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (AB290736)

Blocking / Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-IGF2BP1/IMP1 antibody [EPR26408-18] (ab290736) at 1/1000 dilution

Lane 1:

MYC/DDK-tagged mouse IGF2BP1 full-length recombinant protein 10 ng

Lane 2:

MYC/DDK- tagged mouse IGF2BP2 full-length recombinant protein 10 ng

Lane 3:

His-tagged mouse IGF2BP3 full-length recombinant protein 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 63 kDa

false

Exposure time: 48s