Overview

  • Product name

  • Description

    Rabbit polyclonal to PMP22
  • Host species

    Rabbit
  • Specificity

    ab61220 detects endogenous levels of total PMP22 protein.
  • Tested applications

    Suitable for: ELISA, IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human PMP22 aa 111-160.
    Database link: Q01453

  • Positive control

    • WB: Extracts from MDA-MB-435 cells IHC: Human brain tissue ICC/IF: PC12 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab61220 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
ELISA 1/40000.
IHC-P 1/50 - 1/100.
WB 1/500 - 1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa).
ICC/IF Use a concentration of 1 mg/ml.

Target

  • Function

    Might be involved in growth regulation, and in myelinization in the peripheral nervous system.
  • Involvement in disease

    Defects in PMP22 are the cause of Charcot-Marie-Tooth disease type 1A (CMT1A) [MIM:118220]; also known as hereditary motor and sensory neuropathy IA. CMT1A is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT1 group are characterized by severely reduced nerve conduction velocities (less than 38 m/sec), segmental demyelination and remyelination with onion bulb formations on nerve biopsy, slowly progressive distal muscle atrophy and weakness, absent deep tendon reflexes, and hollow feet. CMT1A inheritance is autosomal dominant.
    Defects in PMP22 are a cause of Dejerine-Sottas syndrome (DSS) [MIM:145900]; also known as Dejerine-Sottas neuropathy (DSN) or hereditary motor and sensory neuropathy III (HMSN3). DSS is a severe degenerating neuropathy of the demyelinating Charcot-Marie-Tooth disease category, with onset by age 2 years. DSS is characterized by motor and sensory neuropathy with very slow nerve conduction velocities, increased cerebrospinal fluid protein concentrations, hypertrophic nerve changes, delayed age of walking as well as areflexia. There are both autosomal dominant and autosomal recessive forms of Dejerine-Sottas syndrome.
    Defects in PMP22 are a cause of hereditary neuropathy with liability to pressure palsies (HNPP) [MIM:162500]; an autosomal dominant disorder characterized by transient episodes of decreased perception or peripheral nerve palsies after slight traction, compression or minor traumas.
    Defects in PMP22 are the cause of Charcot-Marie-Tooth disease type 1E (CMT1E) [MIM:118300]; also known as Charcot-Marie-Tooth disease and deafness autosomal dominant. CMT1E is an autosomal dominant form of Charcot-Marie-Tooth disease characterized by the association of sensorineural hearing loss with peripheral demyelinating neuropathy.
    Defects in PMP22 may be a cause of inflammatory demyelinating polyneuropathy (IDP) [MIM:139393]. IDP is a putative autoimmune disorder presenting in an acute (AIDP) or chronic form (CIDP). The acute form is also known as Guillain-Barre syndrome.
  • Sequence similarities

    Belongs to the PMP-22/EMP/MP20 family.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CMT1A antibody
    • CMT1E antibody
    • DSS antibody
    • GAS-3 antibody
    • GAS3 antibody
    • Growth Arrest Specific 3 antibody
    • Growth arrest-specific protein 3 antibody
    • HMSNIA antibody
    • HNPP antibody
    • MGC20769 antibody
    • Peripheral myelin protein 22 antibody
    • PMP-22 antibody
    • PMP22 antibody
    • PMP22_HUMAN antibody
    • Sp110 antibody
    • Trembler antibody
    see all

Images

  • All lanes : Anti-PMP22 antibody (ab61220) at 1/500 dilution

    Lane 1 : extracts from MDA-MB-435 cells
    Lane 2 : extracts from MDA-MB-435 cells with the immunizing peptide at 5 µg

    Lysates/proteins at 5 µg per lane.

    Predicted band size: 17 kDa
    Observed band size: 19 kDa
    why is the actual band size different from the predicted?

  • Ab61220 at 1/50 dilution staining human brain (Left apical lobe) without (left) and with (right) immunizing peptide; paraffin embedded.
  • ICC/IF image of ab61220 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab61220, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

  • Poitelon Y  et al. A dual role for Integrin a6ß4 in modulating hereditary neuropathy with liability to pressure palsies. J Neurochem N/A:N/A (2018). WB ; Mouse . Read more (PubMed: 29315582) »
  • Bertrand MM  et al. Anatomical basis of the coordination between smooth and striated urethral and anal sphincters: loops of regulation between inferior hypogastric plexus and pudendal nerve. Immuno-histological study with 3D reconstruction. Surg Radiol Anat 38:963-72 (2016). Read more (PubMed: 26952718) »
See all 15 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for your email.

I am surprised that ab126769 failed. This is a rabbit monoclonal antibody which is better than the rabbit polyclonal. Could you please explain what kind of results you are observing? Have you done any troubleshooting?

Thanks! I will be looking forward to hearing from you soon.

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Question
Answer

The antibody ab126769 will be dispatched today.

Many thanks for your cooperation!

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Answer

I have placed an order for you however ab90782 is also out of stock and is expected on 19th September. Please let me know if you can wait for 1 week or else I can send you a in-stock rabbit monoclonal ab126769.

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Answer

I apologize for any inconvenience this may have caused.

The product unfortunately is out of stock at the moment; we are expecting it to be back in stock in month of November. So in order to avoid any further delays I would suggest choosing another product from our catalogue e.g. ab90782 is a good alternative.

Could you let me know if you would like to receive a different product; I will then amend your order for next day delivery?

Thank you very much for your cooperation. I will be looking forward to hearing from you soon.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1152244.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

It will take approximately 5 minutes to complete the questionnaire.
Please fill-in all fields so that we can better assist you
1) Abcam product code
– ab61220
2) Abcam order reference number or product batch number
-Lot GR51089-2
3) Description of the problem
-When using your antibody for western blot we cannot detect any band at the correct size. Instead, we always detect a band at the size of around 75 kDa and when not blocking the membrane with 5% milk we can sometimes detect bands at 15 and 27 kDa.
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…):
-Whole cell lysate from breast cancer MCF7 cells
Lysis buffer
-RIPA
Protease inhibitors:
-Complete without EDTA
Phosphatase inhibitors
-None
Reducing agent
-DTT
Boiling for ≥5 min? yes/no
Protein loaded ug/lane or cells/lane 40-100 µg/lane
Positive control Cells differentiated towards schwann cell like lineage. Don´t know if they should express PMP22, but that was the only neuronal cell lysate that we had.
Negative control
5) Percentage of gel 10%
Type of membrane Tried both PVDF and Hybond C
Protein transfer verified Marker, actin and another protein of interest around 55 kDa has been verified with blottning
Blocking agent and concentration 5% drymilk, 5% BSA or no blocking has been tried.
Blocking time 1 hour
Blocking temperature Room temperature
6) Primary antibody (If more than one was used, describe in “additional notes”) :ab61220
Concentration or dilution 1:500
Diluent buffer 1% drymilk in 1xPBS
Incubation time over night
Incubation temperature: 4°C
7) Secondary antibody:
Species: Donkey
Reacts against: Rabbit
Concentration or dilution 1:5000
Diluent buffer 1% drymilk in 1xPBS
Incubation time 1 hour
Incubation temperature: room temperature
Fluorochrome or enzyme conjugate: HRP
8) Washing after primary and secondary antibodies:
Buffer 1x PBS-T
Number of washes 4x
9)Detection method
10) How many times have you run this staining? 8x
Do you obtain the same results every time? Yes (but different between blocked and not blocked membrane)
What steps have you altered to try and optimize the use of this antibody?
-Amount loaded protein
-Membrane type
-Transfer time
-Blocking type and time
-Diluent buffer for primary antibody (have tried 1% BSA once without any difference from the one diluted in milk).

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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Application
Western blot
Sample
Mouse Tissue lysate - other (mouse sciatic and spinal cord)
Loading amount
10 µg
Specification
mouse sciatic and spinal cord
Gel Running Conditions
Reduced Denaturing (8-12)
Blocking step
Odyssey Infrared Imaging system block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Dr. Gillian Hunter

Verified customer

Submitted Mar 08 2012

Application
Western blot
Sample
Mouse Tissue lysate - whole (Sciatic nerve)
Loading amount
50 µg
Specification
Sciatic nerve
Gel Running Conditions
Reduced Denaturing (4-20% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 19 2009

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