Anti-SOX10 antibody [SOX10/991]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX10 antibody [SOX10/991] (AB218522)

Immunohistochemical analysis of paraffin-embedded Human breast tissue using ab218522 at a 1/1000 dilution followed by a ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).

Nuclear staining in human breast.
The section was incubated with ab218522 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Hematoxylin was used as a counter stain.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SOX10 antibody [SOX10/991] (AB218522)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling SOX10 with ab218522 at 1/50 dilution followed by secondary antibody ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (green).

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (red).

Confocal image showing nuclear staining in A375 cells.
Negative control : MCF7 (PMID : 23799842).
Nuclear DNA was labelled with DAPI (shown in blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SOX10 antibody [SOX10/991] (AB218522)

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized MCF-7(Human breast adenocarcinoma epithelial cell, Left) / A375 (Human malignant melanoma epithelial cell, Right) cells labelling SOX10 with ab218522 at 1/1000 dilution (0.1µg)/Red followed by a Goat Anti-Mouse IgG (Alexa Fluor^® 488, ab150113) at a 1/2000 dilution compared with a Mouse monoclonal IgG (Black) isotype control and unlabelled control Cell without incubation with primary antibody and secondary antibody / Blue.

Negative control : MCF7.(PMID : 23799842)

• IP

Lab

Immunoprecipitation - Anti-SOX10 antibody [SOX10/991] (AB218522)

SOX10 was immunoprecipitated from 0.35 mg A375 (human malignant melanoma epithelial cell) whole cell lysate with ab218522 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab218522 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-SOX10 antibody [SOX10/991] (ab218522) at 1/30 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate (Input) at 10 µg

Lane 2:

A375 whole cell lysate (+)

Lane 3:

Mouse IgG2b instead of ab218522 in A375 whole cell lysate (-)

Secondary

All lanes:

Immunoprecipitation - Anti-mouse IgG for IP (HRP) (<a href='/en-us/products/secondary-antibodies/mouse-igg-for-ip-hrp-ab131368'>ab131368</a>) at 1/1000 dilution

Observed band size: 56-75 kDa

false

Exposure time: 3s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX10 antibody [SOX10/991] (AB218522)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue using ab218522 at a 1/2000 dilution followed by a ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).

Nuclear staining in rat cerebrum.
The section was incubated with ab218522 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Hematoxylin was used as a counter stain.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX10 antibody [SOX10/991] (AB218522)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue using ab218522 at a 1/2000 dilution followed by a ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).

Nuclear staining in mouse cerebrum.
The section was incubated with ab218522 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Hematoxylin was used as a counter stain.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• WB

Lab

Western blot - Anti-SOX10 antibody [SOX10/991] (AB218522)

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

Lysates should be made freshly and used in WB immediately to minimize protein degradation.
Negative control : MCF7 (PMID : 23799842).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 27115022, 29315345 ).

All lanes:

Western blot - Anti-SOX10 antibody [SOX10/991] (ab218522) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 56-75 kDa

false

Exposure time: 26s