Overview

  • Product name
  • Description
    Rabbit polyclonal to CD45
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Pig, Monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1250 to the C-terminus of Human CD45.

    Read Abcam's proprietary immunogen policy (Peptide available as ab17550.)

  • Positive control
    • This antibody gave a positive signal in Jurkat whole cell lysate and Hodgkins lymphoma tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab10559 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
WB 1/500. Detects a band of approximately 155 kDa (predicted molecular weight: 147 kDa).
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.
  • Involvement in disease
    Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
    Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.
  • Sequence similarities
    Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.
  • Domain
    The first PTPase domain interacts with SKAP1.
  • Post-translational
    modifications
    Heavily N- and O-glycosylated.
  • Cellular localization
    Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • Database links
  • Alternative names
    • B220 antibody
    • CD 45 antibody
    • CD45 antibody
    • CD45 antigen antibody
    • CD45R antibody
    • GP180 antibody
    • L-CA antibody
    • LCA antibody
    • Leukocyte common antigen antibody
    • loc antibody
    • Ly-5 antibody
    • LY5 antibody
    • Ly5, homolog of antibody
    • Lyt-4 antibody
    • OTTHUMP00000033813 antibody
    • OTTHUMP00000033816 antibody
    • OTTHUMP00000033817 antibody
    • OTTHUMP00000038574 antibody
    • Protein tyrosine phosphatase receptor type c polypeptide antibody
    • Protein tyrosine phosphatase, receptor type C antibody
    • protein tyrosine phosphatase, receptor type, C antibody
    • Protein tyrosine phosphatase, receptor type, c polypeptide antibody
    • Ptprc antibody
    • PTPRC_HUMAN antibody
    • Receptor-type tyrosine-protein phosphatase C antibody
    • T200 antibody
    • T200 glycoprotein antibody
    • T200 leukocyte common antigen antibody
    see all

Images

  • IHC image of ab10559 staining in human Hodgkins lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10559, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab10559  rabbit polyclonal to CD45  (1/500) with secondary goat anti-rabbit IgG ab6721 (1/5000).

    Expected molecular weight: 147 kDa. WB exposure time: 1min 30sec    

    Lanes 1 to 10: 20µg of cell lysate per lane
    Lanes 6 to 10: ab17550 CD45 blocking peptide used at 1µg/ml

    Lane 1: HeLa Nuclear Extract (ab10559)
    Lane 2: HeLa Whole Cell Lysate (ab10559)
    Lane 3: A431 Whole Cell Lysate (ab10559)
    Lane 4: Jurkat Whole Cell Lysate (ab10559)
    Lane 5: HEK293 Whole Cell Lysate (ab10559)
    Lane 6: HeLa Nuclear Extract (ab10559+ ab17550)
    Lane 7: HeLa Whole Cell Lysate (ab10559+ ab17550)
    Lane 8: A431 Whole Cell Lysate (ab10559+ ab17550)
    Lane 9: Jurkat Whole Cell Lysate (ab10559+ ab17550)
    Lane 10: HEK293 Whole Cell Lysate (ab10559+ ab17550)

    A strong band, slightly higher was seen in Jurkat cell lysate. The band was blocked using the immunising peptide (ab17550). It is likely that the band is CD45.

  • Asynchronous KM-H2 cells were pelleted and labeled by indirect immunofluorescence. Cells were stained with ab10559 (1/200) for 30min at 4'C, washed and then stained with goat anti-rabbit alexafluor 488 (1/200). Forward/Side scatter were used to eliminate cellular debris. The accompanying marker was applied such that only 2% of the IgG control was positive. Based on the accompanying image, approximately 9.62% of cells exhibited positive staining for anti-CD45. Since KM-H2 have low levels of CD45 transcripts, it is expected that they have low levels of CD45 on their surface which is reflected in the ~9% positive. This image is taken from an Abreview.

References

This product has been referenced in:
See all 13 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Answer

Thank you for contacting us.

I am sorry we do not have epitope details of ab106478, at the moment. We will be updating the datasheet when this information will be available.

I can however recommend using antibody ab10559 in IP, although this hasn't been tested however this antibody will work. If the customer is interested in testing this Ab IP, I can happily provide Abreview testing discount. Full details can found by clicking the following link;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

Recommended compatible secondary antibody;

ab97051; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-HRP-ab97051.html

We are giving away free Molecular Weight Marker Samples; please check

https://www.abcam.com/index.html?pageconfig=resource&rid=14769&utm_campaign=Brand.Giveaway&utm_source=Abcam.Brand&utm_medium=Email&utm_term=webbanner-proteinladder&intGoUser=6184200

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Monkey Tissue sections (Spleen)
Specification
Spleen
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 11 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Tonsil)
Specification
Tonsil
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 10 2014

Answer

Thank you for contacting Abcam.
Earlier today we discussed the Human Mesenchymal Stromal Cell Marker panel ab93758. Based on the information on the datasheet, I compiled the following information for you:
1. Positive selection (CD105, CD29, CD44, CD90)
2. Negative selection (CD45)
The stem cell panel ab93758 consists of 5 unconjugated primary antibodies. These antibodies can be used with the secondary antibodies against mouse or rabbit of your choice. You can therefore adapt the fluorochrome choice to the ability of your machine.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for taking the time to contact us with these details. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab10559 is tested and covered by our 6 month guarantee for use in IHC-P and pig samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, in order to investigate further I would appreciate if you are able to confirm some further details:

1. Please confirm the order number and date of purchase.

2. How was the antibody stored?

4. Could you confirm what the problem with the results was? For example, was there no staining, high background or non specific staining? I would appreciate if you are able to send an image which would help us to assess the results.

5. What dilutions of primary antibody have been tried?
Try incubating overnight 4oC, this can sometimes provide more efficient and specific staining.

6. What dilutions of secondary antibody have been tried? Is the current vial of secondary antibody working well with other primary antibodies? Which secondary antibody has been used (supplier and product code?).

7. I can recommend to try including 0.2% Tween 20 detergent in the wash buffer and antibody dilution buffer. This helps to keep the antibody solubilised.

8. What expression level of CD45 are you expecting in Sclera tissue? Which positive controls are being used?

I hope this information is helpful, thank you for your cooperation. Please do not hesitate to contact me again with the further requested details and I hope we can resolve this case for you as soon as possible.

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Question
Answer

Thank you for calling us yesterday and your interest in our products.

As discussed over the phone, the Anti-CD45 antibody (ab10559) has not as yet been tested with mouse samples to ascertain if it is likely to cross react. However, I have now checked the homology of the immunogen used to raise the antibody with the sequence of the mouse protein, and only 9 of the 17 residues used match. I would therefore not expect the antibody to be able to cross react with the mouse protein. However, as we have not tried it ourselves I would not be able to guarantee this.

I have had a look at the other antibodies we have raised to this target that have been shown to work in IHC with paraffin embedded sections. However, I would say that the antibody ab10559 is most likely to be able to differentiate between the human and mouse protein as most of the others have been raised against the full length CD45 protein from cell preparations.

If you would like to purchase ab10559 and use it with yourhuman and mousesamples, we can offer you a testing discount. This offer means that if you purchase ab10559 as normal, test the antibody with yourhuman and mousesamples and let us know of the results throughsubmitting anAbreview (no matter whether positive or negative) you would be eligible for a free primary antibody of your choice from our catalogue (or the equivalent price of ab10559 off any product). More information on this offer can be found here:

www.abcam.com/collaborationdiscount

Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab10559.

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Question
Answer

DISCOUNT CODE: ***
Expiration date: November 24, 2012

This code will give you 1 free primary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for ab40763 in ICC and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

We recommend that alignment should be over 85% to predict that an antibody will detect in a different species. The alignment between the mouse protein and the human immunogen is below the threshold at 80%. I would therefore suggest this antibody would not detect in this species.


However, we do have two antibodies available which will be able to detect in this two applications in mouse:


https://www.abcam.com/index.html?datasheet=10558 (or use the following: https://www.abcam.com/index.html?datasheet=10558).

https://www.abcam.com/index.html?datasheet=23910 (or use the following: https://www.abcam.com/index.html?datasheet=23910).


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry and interest in our products.

I can confirm that this panel contains 5 primary antibodies which recognize human mesenchymal stromal cells: ab6124, ab10559, ab23894, ab44967, ab52971. The datasheets can be found at these sites:

https://www.abcam.com/cd44-antibody-f10-44-2-ab6124.html

https://www.abcam.com/cd45-antibody-ab10559.html

https://www.abcam.com/cd90--thy1-antibody-af-9-ab23894.html

https://www.abcam.com/cd105-antibody-105c02-ab44967.html

https://www.abcam.com/integrin-beta-1-antibody-ep1041y-carboxyterminal-end-ab52971.html

There are some very informative and useful websites summarizing the different CD markers. You may wish to recommend to your customer for further consideration:

https://docs.abcam.com/pdf/immunology/cdantigen_poster.pdf

http://pathologyoutlines.com/cdmarkers.html

If you need any further assistance in the future, please do not hesitate to contact me.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Pig Tissue sections (Epithelial)
Specification
Epithelial
Fixative
Acetone
Permeabilization
No

Mr. Daniel Holod-Betz

Verified customer

Submitted Apr 11 2012

1-10 of 18 Abreviews or Q&A

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