Overview

  • Product name
  • Description
    Rabbit polyclonal to CD41
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic peptide corresponding to Human CD41 (N terminal). Synthetic peptide derived from the N terminal domain of Human CD41.
    Database link: P08514

  • General notes

    This product was changed from whole antiserum to Protein A purified on 15th June 2016. The following lots are from whole antiserum and are still in stock as of 15th June 2016, GR264406-3, GR250036-5 , GR250036-4. Lot numbers higher than GR264406-3, will be Protein A purified. Please note that the dilutions may need to be adjusted accordingly.

Properties

Applications

Our Abpromise guarantee covers the use of ab63983 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 1/100 - 1/5000. Predicted molecular weight: 113 kDa.
ICC/IF Use at an assay dependent concentration. PubMed: 20942599
IHC-P Use at an assay dependent concentration.

Target

  • Function
    Integrin alpha-IIb/beta-3 is a receptor for fibronectin, fibrinogen, plasminogen, prothrombin, thrombospondin and vitronectin. It recognizes the sequence R-G-D in a wide array of ligands. It recognizes the sequence H-H-L-G-G-G-A-K-Q-A-G-D-V in fibrinogen gamma chain. Following activation integrin alpha-IIb/beta-3 brings about platelet/platelet interaction through binding of soluble fibrinogen. This step leads to rapid platelet aggregation which physically plugs ruptured endothelial cell surface.
  • Tissue specificity
    Isoform 1 and isoform 2 were identified in platelets and megakaryocytes, but not in reticulocytes or in Jurkat and U937 white blood cell line. Isoform 3 is expressed by leukemia, prostate adenocarcinoma and melanoma cells but not by platelets or normal prostate or breast epithelial cells.
  • Involvement in disease
    Defects in ITGA2B are a cause of Glanzmann thrombasthenia (GT) [MIM:273800]; also known as thrombasthenia of Glanzmann and Naegeli. GT is the most common inherited disease of platelets. It is an autosomal recessive disorder characterized by mucocutaneous bleeding of mild-to-moderate severity and the inability of this integrin to recognize macromolecular or synthetic peptide ligands. GT has been classified clinically into types I and II. In type I, platelets show absence of the glycoprotein IIb/beta-3 complexes at their surface and lack fibrinogen and clot retraction capability. In type II, the platelets express the glycoprotein IIb/beta-3 complex at reduced levels (5-20% controls), have detectable amounts of fibrinogen, and have low or moderate clot retraction capability. The platelets of GT 'variants' have normal or near normal (60-100%) expression of dysfunctional receptors.
  • Sequence similarities
    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • antigen CD41 antibody
    • BDPLT16 antibody
    • BDPLT2 antibody
    • CD41 antibody
    • CD41B antibody
    • form 2 antibody
    • GP2B antibody
    • GPalpha IIb antibody
    • GPIIb antibody
    • GT antibody
    • GTA antibody
    • HPA3 antibody
    • Integrin alpha 2b antibody
    • Integrin alpha IIb antibody
    • Integrin alpha-IIb light chain antibody
    • Integrin, alpha 2b (platelet glycoprotein IIb of IIb/IIIa complex, antigen CD41) antibody
    • ITA2B_HUMAN antibody
    • Itga2b antibody
    • ITGAB antibody
    • platelet fibrinogen receptor, alpha subunit antibody
    • platelet glycoprotein IIb of IIb/IIIa complex antibody
    • Platelet membrane glycoprotein IIb antibody
    • platelet specific antigen BAK antibody
    • PPP1R93 antibody
    see all

Images

  • All lanes : Anti-CD41 antibody (ab63983) at 1/10000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HepG2 whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 113 kDa
    Observed band size: 113 kDa


    Exposure time: 15 seconds

    See Abreview

  • All lanes : Anti-CD41 antibody (ab63983) at 1/5000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 113 kDa
    Observed band size: 113 kDa


    Exposure time: 10 seconds

    See Abreview

References

This product has been referenced in:
  • Han X  et al. Linarin prevents LPS-induced acute lung injury by suppressing oxidative stress and inflammation via inhibition of TXNIP/NLRP3 and NF-?B pathways. Int J Mol Med 42:1460-1472 (2018). Read more (PubMed: 29845284) »
  • Lee J  et al. Fully Dedifferentiated Chondrocytes Expanded in Specific Mesenchymal Stem Cell Growth Medium with FGF2 Obtains Mesenchymal Stem Cell Phenotype In Vitro but Retains Chondrocyte Phenotype In Vivo. Cell Transplant 26:1673-1687 (2017). Read more (PubMed: 29251111) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 14 Q&A

Answer

Thank you for confirming those details. I have now arranged for a new vial of ab63983 to be sent to you free of charge. This is on the order number 1162584.

Please do not worry about the order number and lot number, I have this information to hand (order number xxxx of the 6th June 2012 with the lot number GR86387-1). It would however be very helpful if you could complete the questionnaire attached.

Please do also let me know how you get on with the new antibody. I look forward to receiving your reply.

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Answer

Thank you for contacting us yesterday and reporting the problems encountered with the Anti-CD41/Integrin alpha 2b antibody (ab63983).

As discussed over the phone, I am able to offer you an alternative lot of ab63983 if you would like to try it. If you would like for me to arrange this please do let me know and could you confirm the following deliver address is correct:

xxxxxx


In order to investigate this problem further, would you mind giving me a few more details of the problems you have been encountering? To this end I have attached a questionnaire that should only take about 5 minutes to complete. This information will be very helpful for us to investigate this problem further and initiate any additional testing where necessary.

I look forward to receiving your reply.

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Answer

Thank you for getting back to me and sorry for the delay in my response.

I have been having some difficulty in finding further choices which may be suitable for your experiments due to the stringent constraints. We have a number of VCAM-1 antibodies, unfortunately none which have been validated in paraffin embedded tissue sections as yet. One such example is the https://www.abcam.com/VCAM1-antibody-324-ab78712.htmlwhich has been used in methanol fixed and frozen sections. It has not been tested with paraffin embedded sections but this does not mean that it would not work, simply that we have not tried it as yet and would not be able to guarantee that is would work.

The only other VCAM-1 antibodies I can suggest it may be worth you having alook at is https://www.abcam.com/VCAM1-antibody-MVCAM-A-429-ab27560.htmland https://www.abcam.com/VCAM1-antibody-M-K-2-ab19569.html(even though they have not as yet been tested with rat samples) as they are raised in rat themselves and would therefore be compatible with the mouse tissue and your other rabbit polyclonal against CD41. I would have a look through the literature to have a look to see if this marker has been used previously to detect the blood vessels as I found some reference to it being more prevalently localised with activated endothelial cells.

I have had another look at the antibodies we have against VE Cadherin and it may be worth trying the mouse monoclonal https://www.abcam.com/VE-Cadherin-antibody-BV9-ab7047.htmlalthough care would have to be taken to reduce the background when performing mouse on mouse staining (details of how to acieve this can be found fromthe link below). Alternatively https://www.abcam.com/VE-Cadherin-antibody-BV13-ab91064.htmlis a rat monoclonal but has as yet not been tested in IHC.

https://www.abcam.com/index.html?pageconfig=resource&rid=11465

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

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Answer

Thank you for contacting us and your interest in our products.

I have tried to find some alternative suggestions for your experiment that you may want to consider. I found no further antibodies against VE-Cadherin that might be suitable and as ab33168 is a rabbit polyclonal, this would be difficult to use in dual staining with ab63983.

One suggestion is goat anti-VEGF receptor 1 (ab10972). This has been tested with both mouse and rat samples and should be compatible with your current antibody you are using.

I am currently looking into whether we have any antibodies which may be suitable for either mouse or rat sample and would be predicted to work in both tissue and I will get back to you with this information as soon as I can. I am sorry for the delay and any inconvenience this is causing you.

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Answer

I am writing to inquire if the heat induced antigen retrieval step has helped to improve the results? I would be pleased to receive an update on how these experiments are progressing.
If the antibody is still not working as stated on the datasheet, we would be happy to replace or refund the antibody.
I wish you the best of luck with your research and I look forward to hearing from you. Please do not hesitate to contact us if you have any further questions.

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Answer

Thank you very much for your answer.
Following your answer on the antigen retrieval, I would like to confirm one more point. Indeed, have the slides been de-paraffinized by sequential incubations? If the slides have not been deparaffinized, this can lead to no staining. As a guideline for deparaffinization, I can recommend also our online protocol under the following link
https://www.abcam.com/index.html?pageconfig=resource&rid=11487
https://www.abcam.com/index.html?pageconfig=resource&rid=11384
In general, when heat induced antigen retrieval is recommended, it is by boiling the samples for a few minutes in a high or low pH buffer. This will allow the protein to unfold and refold in a different manner. Please see also the following publication for more details on antigen retrieval with heat and different buffers:
J Histochem Cytochem. 2005 Nov;53(11):1311-21. Epub 2005 Jul 11.
Please do let me know whether you have any questions, whether the above points are helpful or if you have already performed them.
As mentioned previously, if the antibody is faulty, we will be happy to replace it.
Thank you very much for your cooperation.

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Question

Good morning,
Please find in enclosed (PowerPoint file) our results for these two antibodies, and below the questionnaire.
1) Abcam product code : ab74449 and ab63983

2) Abcam order reference number or product batch number : 1055537
3) Description of the problem : nonspecific staining
4) Sample preparation:
Species : Human
Type of sample : only formalin fixed paraffin embedded sections
Negative control : sample without primary antibody
5) Fixation step :
Yes: formalin 4%
Fixation time : minimum 48h
Fixation temperature : room temperature
6) Antigen retrieval method : heating +60°C
7) Permeabilization method : no permeabilization step
8) Blocking agent : no blocking agent
9) Endogenous peroxidases blocked? Yes
solution from the EnVision + Dual Link System HRP DAB+ (ref: K406511-2) kit form DAKO
Concentration = 0.3% hydrogen peroxide containing sodium azide and levamisole
Blocking time = 10 min
Blocking temperature = room temperature
Endogenous biotins blocked? No
10) Primary antibody :
a) ab74449 :
- ready-to-use (prediluted antibody), 45 minutes at room temperature
- 1:100 in the Antibody Diluent from DAKO (ref: S080981), overnight at +4°C
b) ab63983
- 1:200 in the Antibody Diluent from DAKO (ref: S080981), 45 minutes at room temperature
- 1:1000 in the Antibody Diluent from DAKO (ref: S080981), overnight at +4°C
11) Secondary antibody:
solution from the EnVision + Dual Link System HRP DAB+ (ref: K406511-2) kit form DAKO
= peroxidase labeled polymer conjugated to goat anti-mouse and goat anti-rabbit immunoglobulins in Tris-HCl buffer containing stabilizing protein and antimicrobial agent.
Incubation time : 30 minutes at room temperature
12) Washing after primary and secondary antibodies:
Buffer : PBS 1X
Number of washes : 3 x 5 minutes
13) Detection method : solutions from the EnVision + Dual Link System HRP DAB+ (ref: K406511-2) kit form DAKO
= substrate buffer solution, pH 7.5, containing hydrogen peroxide and a preservative + 3,3'-diaminobenzidine chromogen solution
14) How many times have you run this staining? 4 times
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? The primary antibody Incubation (time and temperature), the primary antibody dilution, and the detecting solution incubation.
Thank you in advance.
Best regards,

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Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that 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.
Having reviewed this case also with the laboratory, I would like to offer some suggestions to help optimize the results from these antibodies. I would also appreciate if you can confirm some further details:
1.) Can you confirm that the negative control, where only the primary antibody is omitted, has no background staining?
Even in this case, I can recommend to include a blocking step for both antibodies. I would recommend to use 3% BSA solution or 10% serum of the species of the secondary antibody.
2.) I agree that the use of the antibody ab74449 neat as well as 1:100 and the ab63983 at the used dilutions should provide good results and can therefore not provide other recommendations in regards to the dilution.
3.) Can you please explain what you mean with the antigen retrieval at 60°C?
Although the laboratory has obtained good staining for theab74449 without antigen retrieval, I can recommend to test different antigen retrieval methods. Indeed, the need of antigen retrieval depends also on the fixation time of the samples. We generally recommend to not fix samples for more than 72 hours, as overfixation of the samples can lead to unsatisfactory results as well.
We recommend normally to perform heat induced antigen retrieval by heating the samples in either citrate buffer pH6 or EDTA buffer pH 9 to 98°C for different times, such as 20 minutes.
4.) Can you please also confirm how the antibodies have been stored?
Please do let me know your comments on the above points.
In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.
I hope this information is helpful, and I thank you for your cooperation.

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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 have currently no reason to think there is a problem with a batch of ab63983. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

Even though you have provided protocol information, I would appreciate if you could provide some more details so I can investigate this issue better. I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Can you please take 5 to 10 minutes to fill the key protocol steps (fixation, antigen retrieval, controls,...) and provide an image of the results? I would also appreciate if you could detail the key steps for both, the frozen section IHC as well as the paraffin embedded IHC.
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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Question
Answer

Thank you for your call today and for your questions. Please see below for information about our testing discount program, and let me know if you have any further questions about these antibodies or our IHC protocols.

Sinceab63983 has not been tested frozen tissue sections (IHC-Fr), I can offer a discount off a future purchase if you buy ab63983 now, test it inIHC-Fr and submit feedback to us in the form of an Abreview. The discount would be worth 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab63983 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab63983 so please wait for my reply before ordering.

2. Purchase ab63983 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-Fr.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab63983 turns out to be unsuitable for IHC-Fr, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us.

I'm sorry to hear that this lot of ab63983 is giving you and your colleagues trouble. We have not had a recent complaint regarding this lot number. However, since it does not appear to be working as expected, I would be happy to send you a replacement vial if you are interested.

In your reply, please include the Abcam order reference number or your PO number so I can process your request.

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1-10 of 14 Q&A

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