Anti-FMRP antibody [EPR23852-90] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

FMRP was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell ) whole cell lysate with ab259335 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259335 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell ) whole cell lysate 10ug

Lane 2 : ab259335 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259335 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>)

Predicted band size: 71 kDa

Observed band size: 80 kDa

false

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse cerebrum tissue sections.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on mouse cerebrum.

Panel B : anti-GPR17 staining oligodendrocytes in mouse cerebrum.

Panel C : anti-P2RY12 staining microglia in mouse cerebrum.

Panel D : anti-FMR1 staining neurons in mouse cerebrum.

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

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution, and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat hippocampus tissue staining TRIM46 with ab307967 at a 1 : 5000 (0.102 ug/ml) dilution, FMRP with ab259335 at 1 : 10000 (0.04 ug/ml) dilution and Serotonin transporter with ab308443 at a 1 : 500 (0.052 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-TRIM46 (green; Opal^™520), anti-FMRP (magenta; Opal^™690) and anti-Serotonin transporter (yellow; Opal^™570) on rat hippocampus.
Panel B : anti-TRIM46 staining the proximal part of the axon in rat hippocampus.
Panel C : anti-FMRP staining neurons in rat hippocampus.
Panel D : anti-Serotonin transporter staining dendrites in rat hippocampus..
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307967, ab259335 and ab308443 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 solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining TRIM46 with ab307967 at a 1 : 5000 (0.102 ug/ml) dilution, FMRP with ab259335 at 1 : 10000 ( 0.04 µg/ml) dilution and Serotonin transporter with ab308443 at a 1 : 500 (0.052 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-TRIM46 (green; Opal^™520), anti-FMRP (magenta; Opal^™690) and anti-Serotonin transporter (yellow; Opal^™570) on mouse hippocampus.
Panel B : anti-TRIM46 staining the proximal part of the axon in mouse hippocampus.
Panel C : anti-FMRP staining neurons in mouse hippocampus.
Panel D : anti-Serotonin transporter staining dendrites in mouse hippocampus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307967, ab259335 and ab308443 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 solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse spinal cord tissue.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on mouse spinal cord.
Panel B : anti-GPR17 staining oligodendrocytes in mouse spinal cord.
Panel C : anti-P2RY12 staining microglia in mouse spinal cord.
Panel D : anti-FMR1 staining neurons in mouse spinal cord.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution, and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue sections.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on mouse hippocampus.

Panel B : anti-GPR17 staining oligodendrocytes in mouse hippocampus.

Panel C : anti-P2RY12 staining microglia in mouse hippocampus.

Panel D : anti-FMR1 staining neurons in mouse hippocampus.

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

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution, and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded rat cerebrum tissue sections.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on rat cerebrum.

Panel B : anti-GPR17 staining oligodendrocytes in rat cerebrum.

Panel C : anti-P2RY12 staining microglia in rat cerebrum.

Panel D : anti-FMR1 staining neurons in rat cerebrum.

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

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution, and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Nuclear counterstaining with DAPI.

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse cerebellum tissue sections.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on mouse cerebellum.

Panel B : anti-GPR17 staining oligodendrocytes in mouse cerebellum.

Panel C : anti-P2RY12 staining microglia in mouse cerebellum.

Panel D : anti-FMR1 staining neurons and Purkinje cells in mouse cerebellum.

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

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded rat hippocampus tissue sections.

Panel A : merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-FMR1 (gray; Opal™570) on rat hippocampus.

Panel B : anti-GPR17 staining oligodendrocytes in rat hippocampus.

Panel C : anti-P2RY12 staining microglia in rat hippocampus.

Panel D : anti-FMR1 staining neurons in rat hippocampus.

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

The section was incubated in three rounds of staining : in the order of ab316105 at a 1/500 dilution, ab300140 at a 1/40000 dilution, and ab259335 at a 1/10000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Nuclear counterstaining with DAPI.

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

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.

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling FMRP with ab259335 at 1/10000 (0.04 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse testis (PMID : 16790844). The section was incubated with ab259335 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized C2C12 (Mouse myoblast) cells labelling FMRP with ab259335 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labelling FMRP with ab259335 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong cytoplasmic and weak nuclear staining in C2C12 cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling FMRP with ab259335 at 1/10000 (0.04 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat testis. The section was incubated with ab259335 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labeling FMRP with ab259335 at 1/10000 (0.04 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining on mouse cardiac muscle (PMID : 7633436). The section was incubated with ab259335 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling FMRP with ab259335 at 1/10000 (0.04 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum. The section was incubated with ab259335 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling FMRP with ab259335 at 1/10000 (0.04 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID : 24463622). The section was incubated with ab259335 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebellum labelling Noradrenaline transporter with ab254361 at 1/100 (B), GPCR GPR17 with ab314307 at 1/2000 dilution (C) and FMRP with ab259335 at 1/10000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and Nuclear DNA was labeled with DAPI (shown in blue). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Noradrenaline transporter (magenta; Opal™690), anti-GPCR GPR17 (green; Opal™520) and anti-FMRP (gray; Opal™570) on mouse cerebellum.

Panel B : anti-Noradrenaline transporter staining nerves in mouse cerebellum.

Panel C : anti-GPCR GPR17 staining oligodendrocytes in mouse cerebellum.

Panel D : anti-FMRP staining neurons in mouse cerebellum.

The section was incubated in three rounds of staining : in the order of ab254361, ab314307 and ab259335 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum labelling Noradrenaline transporter with ab254361 at 1/100 (B), GPCR GPR17 with ab314307 at 1/2000 dilution (C) and FMRP with ab259335 at 1/10000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and Nuclear DNA was labeled with DAPI (shown in blue). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Noradrenaline transporter (magenta; Opal™690), anti-GPCR GPR17 (green; Opal™520) and anti-FMRP (gray; Opal™570) on mouse cerebrum.

Panel B : anti-Noradrenaline transporter staining nerves in mouse cerebrum.

Panel C : anti-GPCR GPR17 staining oligodendrocytes in mouse cerebrum.

Panel D : anti-FMRP staining neurons in mouse cerebrum.

The section was incubated in three rounds of staining : in the order of ab254361, ab314307 and ab259335 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebellum labelling Noradrenaline transporter with ab254361 at 1/100 (B), GPCR GPR17 with ab314307 at 1/2000 dilution (C) and FMRP with ab259335 at 1/10000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and Nuclear DNA was labeled with DAPI (shown in blue). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Noradrenaline transporter (magenta; Opal™690), anti-GPCR GPR17 (green; Opal™520) and anti-FMRP (gray; Opal™570) on rat cerebellum.

Panel B : anti-Noradrenaline transporter staining nerves in rat cerebellum.

Panel C : anti-GPCR GPR17 staining oligodendrocytes in rat cerebellum.

Panel D : anti-FMRP staining neurons in rat cerebellum.

The section was incubated in three rounds of staining : in the order of ab254361, ab314307 and ab259335 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum labelling Noradrenaline transporter with ab254361 at 1/100 (B), GPCR GPR17 with ab314307 at 1/2000 dilution (C) and FMRP with ab259335 at 1/10000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and Nuclear DNA was labeled with DAPI (shown in blue). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Noradrenaline transporter (magenta; Opal™690), anti-GPCR GPR17 (green; Opal™520) and anti-FMRP (gray; Opal™570) on rat cerebrum.
Panel B : anti-Noradrenaline transporter staining nerves in rat cerebrum.
Panel C : anti-GPCR GPR17 staining oligodendrocytes in rat cerebrum.
Panel D : anti-FMRP staining neurons in rat cerebrum.

The section was incubated in three rounds of staining : in the order of ab254361, ab314307 and ab259335 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat cerebrum tissue staining TRIM46 with ab307967 at a 1 : 5000 (0.102 ug/ml) dilution, FMRP with ab259335 at 1 : 10000 (0.04 ug/ml) dilution and Serotonin transporter with ab308443 at a 1 : 500 (0.052 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-TRIM46 (green; Opal^™520), anti-FMRP (magenta; Opal^™690) and anti-Serotonin transporter (yellow; Opal^™570) on rat cerebrum.
Panel B : anti-TRIM46 staining the proximal part of the axon in rat cerebrum.
Panel C : anti-FMRP staining neurons in rat cerebrum.
Panel D : anti-Serotonin transporter staining dendrites in rat cerebrum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307967, ab259335 and ab308443 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 solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse cerebrum tissue staining TRIM46 with ab307967 at a 1 : 5000 (0.102 ug/ml) dilution, FMRP with ab259335 at 1 : 10000 (0.04 ug/ml) dilution and Serotonin transporter with ab308443 at a 1 : 500 (0.052 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-TRIM46 (green; Opal^™520), anti-FMRP (magenta; Opal^™690) and anti-Serotonin transporter (yellow; Opal^™570) on mouse cerebrum.
Panel B : anti-TRIM46 staining the proximal part of the axon in mouse cerebrum.
Panel C : anti-FMRP staining neurons in mouse cerebrum.
Panel D : anti-Serotonin transporter staining dendrites in mouse cerebrum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307967, ab259335 and ab308443 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 solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat spinal cord tissue staining TRIM46 with ab307967 at a 1 : 5000 (0.102 ug/ml) dilution, FMRP with ab259335 at 1 : 10000 (0.04 ug/ml) dilution and Serotonin transporter with ab308443 at a 1 : 500 (0.052 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-TRIM46 (green; Opal^™520), anti-FMRP (magenta; Opal^™690) and anti-Serotonin transporter (yellow; Opal^™570) on rat spinal cord.
Panel B : anti-TRIM46 staining the proximal part of the axon in rat spinal cord.
Panel C : anti-FMRP staining neurons in rat spinal cord.
Panel D : anti-Serotonin transporter staining dendrites in rat spinal cord.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307967, ab259335 and ab308443 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 solution2) for 20 mins

• IP

Unknown

Immunoprecipitation - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

FMRP was immunoprecipitated from 0.35 mg C2C12 (Mouse myoblast) whole cell lysate 10ug with ab259335 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259335 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : C2C12 (Mouse myoblast) whole cell lysate 10ug

Lane 2 : ab259335 IP in C2C12 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259335 in C2C12 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>)

Predicted band size: 71 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using the same antibody clone in a different buffer formulation (ab259335).

Lanes 1 - 4 : Merged signal (red and green). Green - ab259335 observed at 77 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab259335 was shown to react with FMRP in western blot. The band observed in the CRISPR/Cas9 edited lysate lane below 77 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab259335 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

FMRP CRISPR/Cas9 edited HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Human brain cell lysate at 20 µg

Predicted band size: 71 kDa

Observed band size: 77 kDa

false

• WB

Lab

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate 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 1/100000 dilution

Predicted band size: 71 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using the same antibody clone in a different buffer formulation (ab17722).

Western blot : Anti-FMRP antibody ab17722 staining at 1 µg/mL, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in FMR1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FMRP antibody (<a href='/en-us/products/primary-antibodies/fmrp-antibody-ab17722'>ab17722</a>) at 1 µg/mL

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human FMR1 knockout U-87 MG cell line (<a href='/en-us/products/cell-lines/human-fmr1-knockout-u-87-mg-cell-line-ab306664'>ab306664</a>) at 20 µg

Lane 3:

Wild-type A549 at 20 µg

Lane 4:

Western blot - Human FMR1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-fmr1-knockout-a549-cell-line-ab288956'>ab288956</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 71 kDa

Observed band size: 75 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using the same antibody clone in a different buffer formulation (ab109741).

Western blot : Anti-FMRP antibody ab109741 staining at 1 µg/mL, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in FMR1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 Donkey anti-Goat 800CW & Donkey anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FMRP antibody (<a href='/en-us/products/primary-antibodies/fmrp-antibody-ab109741'>ab109741</a>) at 1 µg/mL

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human FMR1 knockout U-87 MG cell line (<a href='/en-us/products/cell-lines/human-fmr1-knockout-u-87-mg-cell-line-ab306664'>ab306664</a>) at 20 µg

Lane 3:

Wild-type A549 at 20 µg

Lane 4:

Western blot - Human FMR1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-fmr1-knockout-a549-cell-line-ab288956'>ab288956</a>) at 20 µg

Secondary

All lanes:

Donkey anti-Goat 800CW & Donkey anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 71 kDa

Observed band size: 75 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Multiple bands could be seen due to alternative splicing of FMRP. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse hippocampus tissue lysate at 20 µg

Lane 3:

Rat hippocampus tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 71 kDa

Observed band size: 75 kDa,80 kDa

false

• WB

Supplier Data

Western blot - Anti-FMRP antibody [EPR23852-90] - BSA and Azide free (AB277489)

This data was developed using ab259335, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 81 seconds.

Negative control : Mouse heart(PMID : 7633436).

Multiple bands could be seen due to alternative splicing of FMRP

All lanes:

Western blot - Anti-FMRP antibody [EPR23852-90] (<a href='/en-us/products/primary-antibodies/fmrp-antibody-epr23852-90-ab259335'>ab259335</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 71 kDa

Observed band size: 75 kDa,80 kDa

false