Overview

  • Product name
    Anti-Thymosin beta 4 antibody
    See all Thymosin beta 4 primary antibodies
  • Description
    Rabbit polyclonal to Thymosin beta 4
  • Host species
    Rabbit
  • Specificity
    Recognises a synthetic Human Thymosin beta 4 peptide(aa 1-43). The antibody has not been tested on the endogenous protein.ab14334 also detects Thymosin beta 9 and Thymosin beta 10 along with Thymosin beta 4.
  • Tested applications
    Suitable for: ELISA, ICC, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Horse, Chicken, Cow
  • Immunogen

    Synthetic peptide: SDKPDMAEIE KFDKSKLKKT ETQEKNPLPS KETIEQEKQA GES conjugated to KLH, corresponding to amino acids 1-43 of Human Thymosin beta 4

  • Positive control
    • Thymosin beta 4 synthetic peptide

Properties

Applications

Our Abpromise guarantee covers the use of ab14334 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 1/4000.
ICC 1/100.
IHC-P 1/100 - 1/1000.
WB Use at an assay dependent concentration. See Smart et al. Non specific bands were reported by the researcher who published this reference.

Target

References

This product has been referenced in:
  • Kannan L  et al. Identification and characterization of thymosin beta-4 in chicken macrophages using whole cell MALDI-TOF. Ann N Y Acad Sci 1112:425-34 (2007). Read more (PubMed: 17947593) »
  • Smart N  et al. Thymosin beta4 induces adult epicardial progenitor mobilization and neovascularization. Nature : (2006). Read more (PubMed: 17108969) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question

Thank you for your reply. I have completed the questionnaire and have also attached pictures. I would be grateful if you could let me know if there are any differences in concentration/activity between the two lots.

Many thanks



Order Details
Antibody code: ab14334

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number GR2453-5

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)

Stored in aliquots at -20C


Description of the problem (high background, wrong band size, more bands, no band etc.)
High background observed when staining section by IHC (DAB)

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Mouse kidney (paraffin – embedded)

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Washes were performed in PBS-T or PBS. Antigen retrieval by incubation with proteinase K (20ug/ml) at 37C for 20mins.

Amount of protein loaded
N/A

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

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Blocking in 10% serum, 1% (w/v) BSA in PBS-T

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
abcam ab14334 rabbit

Diluted in PBS-T with 10% serum and 1% (w/v) BSA 1/200

Incubation overnight at 4C

Washes with PBS-T

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Dako Envision+ kit K4011 – ready made solution – incubation at room temp for 30 mins – washes in PBS

Detection method (ECL, ECLPlus etc.)
DAB

Positive and negative controls used (please specify)
Negative control: no 1ry antibody – showed no staining


Optimization attempts (problem solving)
How many times have you tried the Western?
N/A


Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?
Have not tried altering any steps as antibody from previous lot (944043) was working fine. Would be great to know if the concentration has changed between the lots and need to alter ab dilution.

Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.

Read More
Answer

Thank you for taking the time to complete our questionnaire. 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.


Reviewing this case, I would like to offer some suggestions to help optimize the results between batches. I would also appreciate if you can confirm some further details:

1. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. There have been no reported changes in results. However, there can be some batch variation with polyclonal antibodies. It may be that a lower concentration of antibody is required for this batch to decrease the background signal in your experiments. What dilutions have already been tried?

2. I am sorry to confirm that thi antibody is not tested and covered by the 6 month guarantee in IHC-P, However, as it has worked before for you we can make an exception and provide the guarantee in this case.

3. Antigen retrieval can sometimes require optimization. I can suggest to try different times, for example 5. 10, 20 and 30 minutes. Also, it may be useful to consider trying citrate buffer heat mediated retrieval.

4. I can recommend to include 0.2% Tween in the wash buffer. This will help to wash away ay excess antibody.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this lot of ab14334 Thymosin beta 4 antibody.

I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch.

This antibody is tested and covered by our 6 month guarantee for ELISA and WB and in human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could also provide an image from the batch that has worked, and the one that has, which would help me to assess the results.

Thank you for your time and cooperation. I look forward to receiving the completed questionnaire.



Order Details

Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


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


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


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


Amount of protein loaded


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


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


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


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


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Answer

Thank you for contacting us.

ab14334 is whole serum so it is normal to be brown in colour.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your enquiry. I have been able to source the protocol for direct ELISA which was used with ab14334 and have added it to the datasheet for your convenience. The antibody was tested against the immunizing peptide (Thymosin ß4 synthetic peptide) but you should also see high levels of the protein in thymus extract. Please do not hesitate to contact me if I can be of further help,

Read More

Answer

Thank you for your enquiry regarding ab14334. I can confirm that the antibody has currently only been tested in ELISA on the human synthetic peptide used to generate this antibody. We do not currently supply this peptide or a positive control and I think that human thymus extract may be a good positive control to try. However should you wish to purchased the immunizing peptide please let me know and I will try to source this to add to our catalogue for you. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

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Verified customer

Submitted Aug 09 2005

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