Overview

  • Product name
    Anti-Myelin PLP antibody [plpc 1]
    See all Myelin PLP primary antibodies
  • Description
    Mouse monoclonal [plpc 1] to Myelin PLP
  • Host species
    Mouse
  • Specificity
    This antibody recognizes myelin proteolipid protein
  • Tested applications
    Suitable for: IHC-FoFr, IHC-Fr, WB, ICC/IF, IHC (PFA fixed)more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow
  • Immunogen

    Synthetic peptide:

    C-GRGTKF

    , corresponding to amino acids 272-277 of Myelin PLP.

Properties

Applications

Our Abpromise guarantee covers the use of ab9311 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-FoFr Use at an assay dependent concentration. PubMed: 17626200
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
ICC/IF 1/100.
IHC (PFA fixed) 1/1000.

Target

  • 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.
  • Involvement in disease
    Defects in PLP1 are the cause of leukodystrophy hypomyelinating type 1 (HLD1) [MIM:312080]; also known as Pelizaeus-Merzbacher disease. HLD1 is an X-linked recessive dysmyelinating disorder of the central nervous system in which myelin is not formed properly. It is characterized clinically by nystagmus, spastic quadriplegia, ataxia, and developmental delay.
    Defects in PLP1 are the cause of spastic paraplegia X-linked type 2 (SPG2) [MIM:312920]. SPG2 is characterized by spastic gait and hyperreflexia. In some patients, complicating features include nystagmus, dysarthria, sensory disturbance, mental retardation, optic atrophy.
  • Sequence similarities
    Belongs to the myelin proteolipid protein family.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • HLD1 antibody
    • Lipophilin antibody
    • Major myelin proteolipid protein antibody
    • MMPL antibody
    • Myelin proteolipid protein antibody
    • MYPR_HUMAN antibody
    • PLP 1 antibody
    • PLP antibody
    • PLP/DM20 antibody
    • PLP1 antibody
    • PMD antibody
    • Proteolipid protein 1 antibody
    • SPG2 antibody
    see all

Images

  • ab9311 at a dilution of 1/1000, staining neuronal Myelin PLP  (Alexa 488 secondary at 1/2000) in rat brain tissue (30µm thick coronal sections) on free floating IHC (see protocol link for detailed description). Image taken with a 40x objective. No labeling observed following omission of primary antibody.

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates

  • ab9311 at 1/100 staining rat spinal cord tissue sections by IHC-Fr. The tissue sections were paraformaldehyde fixed and incubated with the antibody for 24 hours. A Texas Red conjugated goat anti-mouse antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Göttle P  et al. Teriflunomide promotes oligodendroglial differentiation and myelination. J Neuroinflammation 15:76 (2018). WB . Read more (PubMed: 29534752) »
  • Pereira GB  et al. Targeted deletion of the antisilencer/enhancer (ASE) element from intron 1 of the myelin proteolipid protein gene (Plp1) in mouse reveals that the element is dispensable for Plp1 expression in brain during development and remyelination. J Neurochem 124:454-65 (2013). WB ; Mouse . Read more (PubMed: 23157328) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Answer

For western blotting we recommend a starting dilution of 1/1000 and that users titrate from there to determine the optimal concentration for their individual experiments.

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Question
Answer

Below please find information regarding your order. Please note it FedEx has delivered this package.
Please feel free to contact us if you have any further questions.
===============================

Items ordered:
1 ab9311 Mouse monoclonal [plpc 1] to Myelin PLP - Oligodendrocyte Marker GR27447
You will be covered by our Abpromise guarantee so long as you are working with the tested applications and species that are listed on each datasheet. Should you have any technical questions please do not hesitate to contact us at: us.technical@abcam.com.
Thank you for buying from Abcam, we wish you success with your future research.

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Question
Answer

Sorry for the confusion regarding this replacement vial. I believe you spoke to my colleague Sherry this morning who informed me she arranged for a replacement vial to be shipped for delivery tomorrow.
If you have any further questions please do not hesitate to contact me.

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Answer

I am arranging the free replacement antibody we discussed last week. I just want to confirm that you wished to receive a new vial of ab9311. If not, please just let me know which antibody you would like and I'll arrange for it to be shipped right away.
Thanks!

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Answer

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.
Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Question

Please read belowthe details of customer's complaint. The images are attached.

Antibody code:
Ms bAb to Myelin PLP [plpc 1] (ab9311)

Antibody storage conditions (temperature/reconstitution etc)

Stock aliquots stored at -20˚C. Dilution for western blot (1:1000 or 1:500 in 4% BSA in TBST with 0.1% Sodium azide) stored at +4˚C.

1:1000 dilution was prepared at arrival of the antibody, before aliquoting and freezing of the stock.

1:500 dilution was prepared from frozen aliquot thawed only at the preparation of the dilution (once).

Description of the problem (high background, wrong band size, more bands, no band etc.)

High background + wrong band size + more bands.

High background and a lot of bands in many sizes. Expected bands are 20 and 26/25 kDa. Only 20 kDa is visible. 26/25 kDa is not visible in most blots or very faint compared to background and other bands.

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)

All samples are from mice cerebellum


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)

1X Sample buffer (final conc. in prepared samples): 88mM Tris-HCL, 2% SDS, 1.7% DTT, Glycerol 11%, Bromophenol Blue.

Protease inhibitors: Complete protease inhibitors cocktail + Pepstatin + Leupeptin.

Samples were heated at 99˚C for 5 min.

Amount of protein loaded

10 - 100ug

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)

Reducing gel, 13% (cerebrum lysate and cerebrum myelin) or 15% (cerebellum lysate) Polyacrylamide.

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)

Transfer buffer: 25mM Tris, 192mM glycine, 20% methanol.

Transfer at 100V, 350mA for 1.5hrs.

Blocking in 4% BSA in TBST for 1hr at room temp.

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

Mouse anti PLP - Ms bAb to Myelin PLP [plpc 1] (Abcam - ab9311).

Dilution for western blot: 1:1000 or 1:500 in 4% BSA in TBST with 0.1% Sodium azide.

Incubation time: Varying – Long incubation (overnight+) at +4˚C, sometimes with additional incubation for 40-100min at room temp.

Wash: 5min in TBST X 3 times.

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

Goat anti Mouse-HRP

Dilution for western blot: 1:10000 in TBST or in 4% Milk in TBST.

Incubation time: 1-2hrs at room temp.

Wash: 5min in TBST X 3 times.

Detection method (ECL, ECLPlus etc.)

ECL

Positive and negative controls used (please specify)

All samples originate in mouse brain tissues, which express PLP protein. Similar samples have been prepared previously and tested successfully with a different anti PLP antibody.

Purified myelin sample is enriched with PLP protein. The same sample was previously tested successfully with a different anti PLP antibody.



Optimization attempts (problem solving)



How many times have you tried the Western?

5 times with alterations.

Have you run a "No Primary" control?

No.

Do you obtain the same results every time? e.g. are the background bands always in the same place?

Background bands are not similar. Some bands appear in all attempts (for example over 15kDa), but others do not (for example (under 37kDa). The relative strength of the bands varies.

What steps have you altered?

Dilution of the primary antibody: 1:1000 and 1:500.

Incubation time with the primary antibody: from 2hrs at RT (no bands at all) to longer incubation times at 4˚C with or without additional incubation 40-100min at RT.

Exposure time: from 1min (1:500) to 1hr (1:1000).

Different samples: Cerebellum P9 lysate, Cerebrum P60 pure myelin, Cerebrum P365 lysate.

Additional Notes

Similar or same samples were prepared previously by me in similar manners (lysis, buffers, amount of protein loaded, sample preparation) and tested successfully with a different anti PLP antibody.

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Answer

Thank you for contacting Abcam regarding ab9311.

I am sorry that the customer is experiencing difficulties with this antibody in WB. I have reviewed the protocol information and images and I agree that this antibody is not working as stated on the datasheet. I would be happy to offer a replacement or credit. Please let me know which is preferred.

I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

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Application
Western blot
Sample
Rabbit Tissue lysate - whole (Brain)
Loading amount
20 µg
Specification
Brain
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-15)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Govindaiah Vinukonda

Verified customer

Submitted Jun 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Oligodendroglia)
Specification
Oligodendroglia
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.01% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 07 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (oligodendroglia)
Loading amount
5 µg
Specification
oligodendroglia
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Aug 30 2007

Answer

Thank you for your enquiry regarding ab9311. This antibody labels myelin PLP protein therefore it will indeed label the myelin rather than the cell body. I would recommend looking at the product ab7474 (in particular the IHC image) which stains for Oligodendrocyte Specific Protein OSP, this appears to label very well the cell bodies of oligodendrocytes. https://www.abcam.com/index.html?datasheet=7474 I hope this information will be useful, please do not hesitate to contact me again if I can be of further assistance,

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1-10 of 15 Abreviews or Q&A

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