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  1. Link

    myelin-plp-antibody-plpc-1-ab9311.pdf

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Anti-Myelin PLP 抗体 [plpc 1] (ab9311)

  • Datasheet
  • SDS
Reviews (5)Q&A (11)References (11)

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Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Western blot - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
  • Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [plpc 1] (ab9311)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [plpc 1] to Myelin PLP
  • Suitable for: IHC, WB, IHC-Fr, ICC/IF
  • Reacts with: Mouse, Rat, Human

こちらの製品もご検討ください

一次抗体
Product image
Anti-BMPR1A antibody (ab174815)
二次抗体
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
タンパク質
Product image
Recombinant Human Myelin PLP protein (His tag) (ab252401)

関連製品

製品の概要

  • 製品名

    Anti-Myelin PLP antibody [plpc 1]
    Myelin PLP 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [plpc 1] to Myelin PLP
  • 由来種

    Mouse
  • 特異性

    This antibody recognizes myelin proteolipid protein
  • アプリケーション

    適用あり: IHC, WB, IHC-Fr, ICC/IFmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide:

    C-GRGTKF

    , corresponding to amino acids 272-277 of Myelin PLP.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 特記事項

    This product was switched from a hybridoma to a recombinant production format on 27th October 2021.

    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 production
    For more information see here.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 精製度

    Protein A purified
  • ポリ/モノ

    モノクローナル
  • クローン名

    plpc 1
  • ミエローマ

    Sp2/0
  • アイソタイプ

    IgG2a
  • 軽鎖の種類

    unknown
  • 研究分野

    • Neuroscience
    • Cell Adhesion Proteins
    • Membrane Proteins
    • Neuroscience
    • Cell Type Marker
    • Glia marker
    • Oligodendrocyte marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Neuroscience Markers
    • Glial
    • Neuroscience
    • Development

関連製品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • Recombinant Protein

    • Recombinant Human Myelin PLP protein (His tag) (ab252401)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab9311の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC
1/5000.
WB (2)
1/1000. Predicted molecular weight: 30 kDa.
IHC-Fr (1)
1/25.
ICC/IF (1)
1/25.
特記事項
IHC
1/5000.
WB
1/1000. Predicted molecular weight: 30 kDa.
IHC-Fr
1/25.
ICC/IF
1/25.

ターゲット情報

  • 機能

    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.
  • 関連疾患

    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.
  • 配列類似性

    Belongs to the myelin proteolipid protein family.
  • 細胞内局在

    Membrane.
  • Target information above from: UniProt accession P60201 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 5354 Human
    • Entrez Gene: 18823 Mouse
    • Entrez Gene: 24943 Rat
    • Omim: 300401 Human
    • SwissProt: P60201 Human
    • SwissProt: P60202 Mouse
    • SwissProt: P60203 Rat
    • Unigene: 1787 Human
    • Unigene: 1268 Mouse
    • Unigene: 4550 Rat
    see all
  • 別名

    • 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

画像

  • Western blot - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Western blot - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    All lanes : Anti-Myelin PLP antibody [plpc 1] (ab9311) at 1/1000 dilution

    Lane 1 : Human cerebellum tissue lysate at 20 µg with NFDM/TBST
    Lane 2 : Human liver tissue lysate at 40 µg with NFDM/TBST

    Blocking peptides at 5 % per lane.

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

    Predicted band size: 30 kDa
    Observed band size: 20.23 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Samples are non-boiled as boiling may cause protein aggregates.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 9247276).

    Negative control: liver (PMID: 2414013).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling Myelin PLP with ab9311 at 1/5000 dilution, followed by secondary LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use. Positive staining on human cerebrum is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use.

    The section was incubated with ab9311 for 30 mins at room temperature and followed by mouse specific IgG antibody for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling Myelin PLP with ab9311 at 1/5000 dilution, followed by secondary LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use. Positive staining on human cerebrum is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use.

    The section was incubated with ab9311 for 30 mins at room temperature and followed by mouse specific IgG antibody for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling Myelin PLP with ab9311 at 1/5000 dilution, followed by secondary LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use. Positive staining on human cerebrum is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection), ready to use.

    The section was incubated with ab9311 for 30 mins at room temperature and followed by mouse specific IgG antibody for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling Myelin PLP with ab9311 at 1:25 (32.36 μg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) secondary at 1:1000 dilution. The nuclear counterstain was 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 was ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1:1000 dilution.

  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat cerebellum tissue labeling Myelin PLP with ab9311 at 1:25 (32.36 μg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) secondary at 1:1000 dilution. The nuclear counterstain was 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 was ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1:1000 dilution.

  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling Myelin PLP with ab9311 at 1:25 (32.36 μg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) secondary at 1:1000 dilution. The nuclear counterstain was 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 was ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1:1000 dilution.

  • Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunohistochemistry (Frozen sections) - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat cerebellum tissue labeling Myelin PLP with ab9311 at 1:25 (32.36 μg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) secondary at 1:1000 dilution. The nuclear counterstain was 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 was ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1:1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [plpc 1] (ab9311)
    Immunocytochemistry/ Immunofluorescence - Anti-Myelin PLP antibody [plpc 1] (ab9311)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling Myelin PLP with ab9311 at 1/25 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1:1000 dilution (Green). is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1:1000 dilution.

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (11)

ab9311 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab9311 は 11 報の論文で使用されています。

  • Cerina M  et al. Myelination- and immune-mediated MR-based brain network correlates. J Neuroinflammation 17:186 (2020). PubMed: 32532336
  • Göttle P  et al. Teriflunomide promotes oligodendroglial differentiation and myelination. J Neuroinflammation 15:76 (2018). WB ; Rat . PubMed: 29534752
  • Narayanan V  et al. Impairment of frequency-specific responses associated with altered electrical activity patterns in auditory thalamus following focal and general demyelination. Exp Neurol 309:54-66 (2018). PubMed: 30048715
  • 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 . PubMed: 23157328
  • Clark K  et al. Gait abnormalities and progressive myelin degeneration in a new murine model of Pelizaeus-Merzbacher disease with tandem genomic duplication. J Neurosci 33:11788-99 (2013). PubMed: 23864668
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-10 of 16 Abreviews or Q&A

Immunocytochemistry abreview for Anti-Myelin PLP antibody [plpc 1]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cultured Cells (NSC-34)
Permeabilization
Yes - 0.05% tritonX
Specification
NSC-34
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

投稿 Dec 28 2019

Western blot abreview for Anti-Myelin PLP antibody [plpc 1] - Oligodendrocyte Marker

Average
Abreviews
Abreviews
abreview image
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
Read More

DR. Govindaiah Vinukonda

Verified customer

投稿 Jun 01 2010

Immunocytochemistry/ Immunofluorescence abreview for Anti-Myelin PLP antibody [plpc 1] - Oligodendrocyte Marker

Average
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Sep 07 2007

Western blot abreview for Anti-Myelin PLP antibody [plpc 1] - Oligodendrocyte Marker

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Aug 30 2007

Question

I would like to know the dilution for ab9311 to be used for western blot.
Could you please email the information by today.

Read More

Abcam community

Verified customer

Asked on Apr 10 2014

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.

Read More

Abcam Scientific Support

Answered on Apr 10 2014

Question

please confirm that the delivery is today.

Read More

Abcam community

Verified customer

Asked on Sep 12 2012

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.

Read More

Abcam Scientific Support

Answered on Sep 12 2012

Question

Please email the delivery date of the free replacement antibody.

Read More

Abcam community

Verified customer

Asked on Sep 11 2012

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.

Read More

Abcam Scientific Support

Answered on Sep 11 2012

Question

ab9311 is not working as expected in IHC-Fr staining of mouse brain sections; primary antibody at 1:100; secondary is texas red conjugate

Read More

Abcam community

Verified customer

Asked on Sep 10 2012

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!

Read More

Abcam Scientific Support

Answered on Sep 10 2012

Question

Thank you for the reply. The customer prefers a credit. Thanks in advance for your assistance.

Read More

Abcam community

Verified customer

Asked on Jan 31 2012

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

Read More

Abcam Scientific Support

Answered on Jan 31 2012

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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Abcam community

Verified customer

Asked on Jan 25 2012

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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Abcam Scientific Support

Answered on Jan 25 2012

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