Anti-c-Fos antibody (ab62147)

IHC-FoFr(untested) with rat brain
sees no staining
Ab: 1/500, 1/1000, 1/5000 for 48 h at 4 degree
secondary Ab seems OK
block with 1% BSA and triton
send testing discount code

ANSWER:

Thank you for your phone call. I am sorry that the antibody is not working well in IHC-FoFr, but since this application has not been tested, I am sending you a testing discount code as compensation.

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab62147 in IHC-FoFr. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-FoFr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. 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: http://www.abcam.com/collaborationdiscount.

send antigen retrieval protocol
trying IHC-FoFr and IHC-Fr on mouse tissue

ANSWER:

Thank you for contacting us.

Here is part of the IHC-P protocol with the antigen retrieval step:

Step 1: De-paraffinisation
Xylene (2 x 10 min), 100% ethanol (2 x 5 min), 95% , 70% ethanol, dH2O (1 x 5 min each)

Step 2: Antigen retrieval (in microwave using antigen retrieval tank)
1mM EDTA pH. 8.0
Pour 1mM EDTA into the tank. Boil (3 min – high power)
Place slides in the tank (5 min – med/low power)
Remove tank from microwave. Top up with 1mM EDTA (5 min med/low power)
Remove tank from microwave. Allow to cool at room temperature (20 min) & transfer to 1x TBS-azide

Step 3: Elimination of endogenous peroxidase activity
1% H2O2, 50% Methanol in dH2O (15 ml 100% Methanol + 15 ml dH2O + 1ml 30% H2O2), a few drops/slide (10 min) in the humidifying chamber.

Step 4: Flush and Wash
Flush slides with excess 1x TBS-azide. Place slides in 1x TBS-azide (5 min)

Step 5: Blocking non-specific binding sites
20% NHS (without heat inactivation) in 1xTBS-azide, 250 ul/slide (1 h) in the humidifying chamber

Step 6: Primary antibody
Diluted in 1% NHS - TBS-azide, 250 ul/slide (overnight) in the humidifying chamber (pour some water at the bottom of chamber to avoid drying of slides)

I realized that I had sent you a testing discount lasttime you called for trying the antibody in IHC-FoFr. But I use an incorrect email address and you most likely did not receivemy email. I will resend it to you. I apologize about this oversighton my end.

Please let me know if the IHC-Fr version will work for your sections. If not, we can discuss replacing, crediting or refunding the antibody.

I wish you good luck with everything.
Please do not hesitate to contact us if you need any more advice or information.

IHC-FoFr(untested) with rat brain
sees no staining
Ab: 1/500, 1/1000, 1/5000 for 48 h at 4 degree
secondary Ab seems OK
block with 1% BSA and triton
send testing discount code

ANSWER:

Thank you for your phone call. I am sorry that the antibody is not working well in IHC-FoFr, but since this application has not been tested, I am sending you a testing discount code as compensation.
DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab62147 in IHC-FoFr. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-FoFr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. 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: http://www.abcam.com/collaborationdiscount.

Dear Sir/Madam, I am looking for an anti-c-Fos antibody for use in double-labelling experiments (immunohistochemistry) on perfusion-fixed, frozen rat brain sections. The antibody should NOT be produced in rabbit. Looking at the Abcam catalogue I found these possibilities: ab87655   Anti-c-Fos antibody (goat) ab62147   Anti-c-Fos antibody (sheep) Which antibody would you suggest for our application? Many thanks,

ANSWER:

Thank you for contacting us with your question. Both of these antibodies has been validated in rat, and they have similar properties so it is difficult to say which would be better for your staining. We haven't tested the antibodies on perfusion-fixed frozen sections (we call this "IHC-FoFr"), so we can not say that either one will be suitable on this type of tissue. However, we do have a testing discount program if you are interested in trying either of them in IHC-FoFr. I hope this information will be useful, but please let me know if you have any further questions or if you would like information about our testing discount program.