Anti-Myelin PLP antibody [EPR23504-106] (ab254363)

Anti-Myelin PLP antibody [EPR23504-106] (ab254363) is a rabbit monoclonal antibody detecting Myelin PLP in Western Blot, IP, IHC-P, IHC-Fr, ICC/IF, mIHC. Suitable for Human, Mouse, Rat.

- Multiplex IHC validated on the Leica BOND® MAX using Opal reagents

- Biophysical QC for unrivalled batch-batch consistency

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Related conjugates and formulations

ab275751
Carrier free
Anti-Myelin PLP antibody [EPR23504-106] - BSA and Azide free
ab313220
Conjugated
Alexa Fluor® 555 Anti-Myelin PLP antibody [EPR23504-106]
ab311737
Conjugated
Alexa Fluor® 594 Anti-Myelin PLP antibody [EPR23504-106]
ab313016
Conjugated
Alexa Fluor® 568 Anti-Myelin PLP antibody [EPR23504-106]
ab319182
Conjugated
Alexa Fluor® 488 Anti-Myelin PLP antibody [EPR23504-106]
ab321506
Conjugated
Alexa Fluor® 750 Anti-Myelin PLP antibody [EPR23504-106]

Images

Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [EPR23504-106] (AB254363), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	WB	mIHC	IHC-Fr	IHC-P
Human	Expected	Expected	Tested	Tested	Expected	Tested
Mouse	Tested	Tested	Tested	Expected	Tested	Tested
Rat	Tested	Expected	Tested	Expected	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes -
Species Rat	Dilution info 1/2000	Notes -
Species Human	Dilution info 1/2000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Species Rat	Dilution info 1/500	Notes Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information PLP1

Function

This is the major myelin protein from the central nervous system. It plays an important role in the formation or maintenance of the multilamellar structure of myelin.

Alternative names

PLP, PLP1, Myelin proteolipid protein, Lipophilin

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23504-106
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-Myelin PLP antibody [EPR23504-106] (ab254363) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunohistochemistry (IHC-Fr), Immunocytochemistry/immunofluorescence (ICC/IF), Multiplex IHC (mIHC) in Human, Mouse, Rat samples.

What is the molecular weight of Myelin PLP?

Anti-Myelin PLP [EPR23504-106] (ab254363) specifically detects a band for Myelin PLP (UniProt: P60201) at a molecular weight of 30kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR23504-106] also available for your convenience:
ab254363, Carrier free - ab275751, Alexa Fluor® 594 - ab311737, Alexa Fluor® 568 - ab313016, Alexa Fluor® 555 - ab313220, Alexa Fluor® 488 - ab319182, Alexa Fluor® 750 - ab321506

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Myelin proteolipid protein or PLP is an integral membrane protein with important roles in the central nervous system. It has an alternate name lipophilin and its molecular weight is approximately 25 kDa. PLP is most abundant in myelin the protective sheath covering nerve fibers and is expressed in oligodendrocytes and Schwann cells. It represents one of the major protein components of central nervous system myelin and facilitates the compact structure of myelin via its structural properties.

Biological function summary

The myelin proteolipid protein contributes to the stability and functionality of the myelin sheath impacting nervous impulse conduction. It plays a structural role in the formation and maintenance of myelin which is essential for rapid signal transmission in the nervous system. PLP can form a complex with DM20 a related isoform which aids in the regulatory processes of myelination. The protein is critical for the compaction and shear resistance of the myelin sheath ensuring efficient neuronal communication.

Pathways

The myelin proteolipid protein is involved in myelination and cell adhesion processes that are part of central nervous system development. It participates in the myelin assembly pathway and interacts with other key proteins like 2'3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) to modulate myelin biogenesis and homeostasis. This interaction is important for the formation and repair of myelin ensuring that signal transduction remains unobstructed in the nervous system.

Associated diseases and disorders

Mutations or misregulations of the myelin proteolipid protein are linked to conditions such as Pelizaeus-Merzbacher disease (PMD) and multiple sclerosis (MS). In PMD altered PLP expression leads to abnormal myelin formation resulting in severe neurological dysfunction. In cases of MS the autoimmune response may involve components of the myelin such as PLP initiating demyelination and impairing nerve conductivity. Other proteins connected to these disorders include myelin basic protein (MBP) which alongside PLP forms the bulk of myelin structure and both are targets in autoimmune attack scenarios.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

16 product images

Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunocytochemistry analysis of rat primary neural/glia cells labelling Myelin PLP with ab254363 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-Myelin Basic Protein rat monoclonal antibody at 1/100 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Confocal image showing cytoplasmic staining in rat primary glia cells.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Myelin PLP with ab254363 at 1/2000 (0.261 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on mouse cerebrum (PMID: 28066178). Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 9247276).
Negative control: liver (PMID: 2414013).
Exposure time: 37 seconds.
All lanes: Western blot - Anti-Myelin PLP antibody [EPR23504-106] (ab254363) at 1/2000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Lane 3: Rat brain tissue lysate at 20 µg
Lane 4: Rat liver tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 30 kDa
Observed band size: 20 kDa, 23 kDa
Immunoprecipitation - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Myelin PLP was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab254363 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254363 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain tissue lysate 5 ug
Lane 2: ab254363 IP in Mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254363 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 6 seconds.
All lanes: Immunoprecipitation - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Predicted band size: 30 kDa
Observed band size: 20 kDa, 23 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunocytochemistry analysis of mouse primary neural/glia cells labelling Myelin PLP with ab254363 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-MAP2 mouse monoclonal antibody (Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267) at 1/200 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Confocal image showing cytoplasmic staining in mouse primary glia cells.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Western blot - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 9247276)
Exposure time: 1 second.
All lanes: Western blot - Anti-Myelin PLP antibody [EPR23504-106] (ab254363) at 1/5000 dilution
All lanes: Human brain tissue lysate at 10 µg
Secondary
All lanes: VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Predicted band size: 30 kDa
Observed band size: 20 kDa, 23 kDa
Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling Myelin PLP with ab254363 at 1/500 (1.042 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebellum. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum tissue labeling Myelin PLP with ab254363 at 1/500 (1.042 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat cerebellum. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Myelin PLP with ab254363 at 1/2000 (0.261 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on rat cerebrum. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling Myelin PLP with ab254363 at 1/500 (1.042 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling Myelin PLP with ab254363 at 1/500 (1.042 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Myelin PLP with ab254363 at 1/2000 (0.261 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human cerebrum (PMID: 29081415). Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Multiplex immunohistochemistry - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Myelin PLP Multiplex immunohistochemistry staining of Human Cerebrum Tissue using rabbit Anti-Myelin PLP antibody
Fluorescence multiplex immunohistochemical analysis of human cerebrum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-MAP2 (Anti-MAP2 antibody [EPR22641-106] - Neuronal Marker ab254263, red; Opal™570) on human cerebrum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and Anti-MAP2 antibody [EPR22641-106] - Neuronal Marker ab254263 (1/4000 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 solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

Image acquisition was performed with Leica SP8 confocal microscope.
Multiplex immunohistochemistry - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Myelin PLP Multiplex immunohistochemistry staining of Human cerebellum tissue using rabbit Anti-Myelin PLP antibody
Fluorescence multiplex immunohistochemical analysis of the human cerebellum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-P2Y12 (Anti-P2Y12 antibody [EPR23511-72] ab254347 red; Opal™570) on human cerebellum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and Anti-P2Y12 antibody [EPR23511-72] ab254347 (1/1000 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. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.
Multiplex immunohistochemistry - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Myelin PLP Multiplex immunohistochemistry staining of Human cerebrum tissue using rabbit Anti-Myelin PLP antibody
Fluorescence multiplex immunohistochemical analysis of the human cerebrum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-P2Y12 (Anti-P2Y12 antibody [EPR23511-72] ab254347, red; Opal™570) on human cerebrum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and Anti-P2Y12 antibody [EPR23511-72] ab254347 (1/1000 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. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.
Multiplex immunohistochemistry - Anti-Myelin PLP antibody [EPR23504-106] (ab254363)
Myelin PLP Multiplex immunohistochemistry staining of Human cerebellum tissue using rabbit Anti-Myelin PLP antibody
Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-MAP2 (Anti-MAP2 antibody [EPR22641-106] - Neuronal Marker ab254263, red; Opal™570) on human cerebellum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and Anti-MAP2 antibody [EPR22641-106] - Neuronal Marker ab254263 (1/4000 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 solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.