重组Anti-Peripherin抗体[EPR23445-28]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling Peripherin with ab246502 at 1/50 dilution (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Peripherin with ab246502 at 1/2000 dilution (0.227 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the ganglion cells in human colon (PMID : 26469323). The section was incubated with ab246502 for 10 mins at room temperature. The immunostaining staining 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labeling Peripherin with ab246502 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line. Tubulin is detected using ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red). The nuclear counter stain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit at 1/1000 dilution.

• IP

Unknown

Immunoprecipitation - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Peripherin was immunoprecipitated from 0.35 mg Human spinal cord lysate with ab246502 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246502 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human spinal cord lysate 10ug

Lane 2 : ab246502 IP in Human spinal cord lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246502 in Human spinal cord lysate

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

Exposure time : 40 seconds.

The 58KD isoform of peripherin observed is consistent with what has been described in the literature (PMID : 20587592, 18287500, 17475883).

All lanes:

Immunoprecipitation - Anti-Peripherin antibody [EPR23445-28] (ab246502)

Predicted band size: 53 kDa

Observed band size: 58 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Peripherin with ab246502 at 1/10,000 dilution (0.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the ganglion cells in rat colon. The section was incubated with ab246502 for 10 mins at room temperature. The immunostaining staining 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Peripherin with ab246502 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining on the ganglion cells in mouse colon is observed. The nuclear counter stain is DAPI (Blue).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Peripherin with ab246502 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining on the ganglion cells in rat colon is observed. The nuclear counter stain is DAPI (Blue).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Peripherin with ab246502 at 1/50 dilution (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunofluorescent analysis of4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling Peripherin with ab246502 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line. Tubulin is detected using ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red). The nuclear counter stain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Peripherin with ab246502 at 1/10,000 dilution (0.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the ganglion cells in mouse colon. The section was incubated with ab246502 for 10 mins at room temperature. The immunostaining staining 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• WB

Unknown

Western blot - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 37 seconds.

The 58KD, 56KD, and 45KD isoforms of peripherin observed are consistent with what has been described in the literature (PMID : 20587592, 18287500, 17475883).

Negative control : Kidney (PMID : 15132161).

All lanes:

Western blot - Anti-Peripherin antibody [EPR23445-28] (ab246502) at 1/1000 dilution

Lane 1:

Human spinal cord lysate at 20 µg

Lane 2:

Human lower limb nerve lysate at 20 µg

Lane 3:

Human nerve lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Human kidney lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) or VeriBlot for IP secondary antibody (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 45 kDa,56 kDa,58 kDa

false

• WB

Unknown

Western blot - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 6 seconds.

The 58KD, 56KD and 45KD isoforms of peripherin observed are consistent with what has been described in the literature (PMID : 20587592, 18287500, 17475883).

All lanes:

Western blot - Anti-Peripherin antibody [EPR23445-28] (ab246502) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 10 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: 53 kDa

Observed band size: 45 kDa,56 kDa,58 kDa

false

• WB

Unknown

Western blot - Anti-Peripherin antibody [EPR23445-28] (AB246502)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 26 seconds.

The 58KD, 56KD, 45KD and 28KD isoforms of peripherin observed are consistent with what has been described in the literature (PMID : 20587592,18287500,17475883).

All lanes:

Western blot - Anti-Peripherin antibody [EPR23445-28] (ab246502) at 1/1000 dilution

Lane 1:

Mouse dorsal ganglion lysate at 20 µg

Lane 2:

Rat dorsal ganglion lysate at 20 µg

Lane 3:

Mouse spinal cord lysate at 20 µg

Lane 4:

Rat spinal cord lysate at 20 µg

Lane 5:

Mouse sciatic nerve 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: 53 kDa

Observed band size: 28 kDa,45 kDa,56 kDa,58 kDa

false