Anti-HA tag antibody [12CA5] (ab16918)

is the light chain lambda or kappa?

I am looking to order some light chain specific secondary for this antibody. Can you please tell me in the HA-antibody is light chain or heavy chain specific?

ANSWER:

Thank you for contacting us.

The lab let me know the following:

We really do not know, as we have never tested for it. Our guess would be kappa, since kappa outnumbers lambda about 20:1 But that is only an educated guess.

We do carry anti-kappa light chain antibodies that react with mouse: ab103827, ab46527 (HRP-conjugated), and ab11662.

http://www.abcam.com/Kappa-light-chain-antibody-ab103827.html
http://www.abcam.com/Kappa-light-chain-antibody-HRP-ab46527.html
http://www.abcam.com/Kappa-light-chain-antibody-MRC-OX-20-ab11662.html

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

Dear Dr.
 
We received a replacement anti-HA antibody (12CA5) from you. The lot number is GR67085-1.
We used the same protocol as described before. For result-3, the samples are ARCaPE and ARCaPE.HA.Sox4 stable cells. The HA.Sox4 expression has been verified and the results have been shown in my previous e-mails. For result-4, the samples are LNCaP and LNCaP.HoxC6.HA. The expression of HA-tagged HOXC6 has been verified using our control antibody [anti-HA antibody from Covance (16B12, #MMS-101R)].
Unfortunately, we could not detect HA-tagged HOXC6 expression in our stable cell line using your antibody. Your antibody can only recognize a non-specific 75 kD protein. If you have any suggestion, please let us know. Otherwise, please refund us because we received two vials of this antibody with two different lot numbers and both of them failed, and we do not want to put further effort.
Thank you.
 
Best Regards,
 
 
Zhongming
 
 
From: technical@abcam.com [technical@abcam.com]
Sent: Tuesday, March 06, 2012 7:03 PM
To: Mou, Zhongming
Subject: Reply from Abcam to your enquiry regarding ab16918 [CCE3574245]
 
 

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Dear Sir/Madam
 
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 1047887.

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.

 

Help us improve our service.
http://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3574245
Best regards,
Keith

Keith Beadle
Scientific Support Specialist
Abcam Inc.
www.abcam.com

Your original inquiry to Abcam:
 
 
 
 
 
Hi Keith,
 
I did the western according to your suggestions. I increased the 1' antibody concention to 1:500 and 1:250, respectively.
The other steps are the same as my previous description. It seems your antibody recognizes a non-specific 75 kD protein. My positive control antibody (anti-HA, clone 16B12, from  Covance) can detect a specific protein. I attached the results.
If we are eligible for a replacement antibody, and its lot number is different from the one I used, please give us a replacement. (The lot number of the anti-HA antibody I used is GR55658-2).
If you do not have this antibody of different lot number, and if we are eligible for a refund, please refund us.
Thank you.
 
Best Regards,
 
Zhongming
 
 
 
From: technical@abcam.com [technical@abcam.com]
Sent: Thursday, March 01, 2012 4:52 PM
To: Mou, Zhongming
Subject: Reply from Abcam to your enquiry regarding ab16918 [CCE3561387]
 
 

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Dear Sir/Madam
 
Thank you for contacting us.

I am indeed sorry that this product has not been performing as expected in your assay. I would also like to thank you for sending me the details of your protocol and images. It is often the case that each antibody will need to be optimized in the lab. I noted that you have used this prodcut at the suggested 1:1000 and at a lower dilution. As this did not result in satisfactory performance may I suggest that you try to increase the amount of antibody that you use?
The antibody is covered under our Abpromise for six months and is guaranteed to work in WB. If you are experiencing issues with this product and we cannot resolve the issue you are having, then I would be happy to either send a replacement antibody or to process a refund.  

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

Help us improve our service.
http://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3561387
Best regards,
Keith

Keith Beadle
Scientific Support Specialist
Abcam Inc.
www.abcam.com

Your original inquiry to Abcam:
 
 
 
 
 
 

 
 
 
 
Dear Ms./Mr.,
 
About two weeks ago, we purchased an anti-HA antibody (12CA5) from you. The catalog number is ab16918. The lot number is GR55658-2.
Our shipping address is the following
Soma Sannigrahi
Whitehead Building Rm 139
Emory University
615 Michael st
Atlanta, GA 30322
 
I used this antibody to detect the expression HA-Sox4 in ARCaPE.HA-Sox4 stable cell line. I also used another anti-HA antibody from Covance (16B12, #MMS-101R) as a control. The 16B12 anti-HA antibody from Covance worked very well. Unfortunately, your antibody (12CA5) did not work. I attached a file of my immunoblot with this email.
The following is my protocol
1. Cell lysates from ARCaPE and ARCaPE.HA.Sox4 stable cell lines were run in SDS-PAGE and transferred to nitrocellulose membrane.
2. For figure 1, membrane was blocked with 1% BSA for 1 hour and incubated with anti-HA (12CA5) at 4 degree overnight at 1:2000 dilution in 1% BSA.
    For figure 3, membrane was blocked with 1% BSA for 1 hour and incubated overnight with a positive control antibody, anti-HA (16B12) at 1 ug/ml.
    For figure 2 and 4, the membrane was blocked with LiCor Odyssey blocking buffer (#927-40000) and incubated at 4 degree overnight with anti-HA (12CA5) at 1:1000 and anti-HA (16B12) at 1 ug/ml, respectively.
3. The next day, membranes were washed and incubated with secondary antibody at room temperature for 1 hour. The secondary antibody is LiCor IRDye® 800CW Conjugated Goat Anti-Mouse IgG (H+L) (#926-32210).
4. Membranes were washed and scanned using Odyssey imaging system.
 
Do you have any suggestions? Is it possible to get a replacement antibody from you with a different lot number?
Thank you.
 
Best Regards,
 
Zhongming Mou
 




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ANSWER:

Thank you for contacting us.

Your credit note ID

I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

I did the western according to your suggestions. I increased the 1' antibody concention to 1:500 and 1:250, respectively. The other steps are the same as my previous description. It seems your antibody recognizes a non-specific 75 kD protein. My positive control antibody (anti-HA, clone 16B12, from  Covance) can detect a specific protein. I attached the results. If we are eligible for a replacement antibody, and its lot number is different from the one I used, please give us a replacement. (The lot number of the anti-HA antibody I used is GR55658-2). If you do not have this antibody of different lot number, and if we are eligible for a refund, please refund us. Thank you.

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 1047887.


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.

I used this antibody to detect the expression HA-Sox4 in ARCaPE.HA-Sox4 stable cell line. I also used another anti-HA antibody from Covance (16B12, #MMS-101R) as a control. The 16B12 anti-HA antibody from Covance worked very well. Unfortunately, your antibody (12CA5) did not work. I attached a file of my immunoblot with this email. The following is my protocol 1. Cell lysates from ARCaPE and ARCaPE.HA.Sox4 stable cell lines were run in SDS-PAGE and transferred to nitrocellulose membrane. 2. For figure 1, membrane was blocked with 1% BSA for 1 hour and incubated with anti-HA (12CA5) at 4 degree overnight at 1:2000 dilution in 1% BSA.     For figure 3, membrane was blocked with 1% BSA for 1 hour and incubated overnight with a positive control antibody, anti-HA (16B12) at 1 ug/ml.     For figure 2 and 4, the membrane was blocked with LiCor Odyssey blocking buffer (#927-40000) and incubated at 4 degree overnight with anti-HA (12CA5) at 1:1000 and anti-HA (16B12) at 1 ug/ml, respectively. 3. The next day, membranes were washed and incubated with secondary antibody at room temperature for 1 hour. The secondary antibody is LiCor IRDye® 800CW Conjugated Goat Anti-Mouse IgG (H+L) (#926-32210). 4. Membranes were washed and scanned using Odyssey imaging system.   Do you have any suggestions? Is it possible to get a replacement antibody from you with a different lot number? Thank you.  

ANSWER:

Thank you for contacting us.

I am indeed sorry that this product has not been performing as expected in your assay. I would also like to thank you for sending me the details of your protocol and images. It is often the case that each antibody will need to be optimized in the lab. I noted that you have used this prodcut at the suggested 1:1000 and at a lower dilution. As this did not result in satisfactory performance may I suggest that you try to increase the amount of antibody that you use?

The antibody is covered under our Abpromise for six months and is guaranteed to work in WB. If you are experiencing issues with this product and we cannot resolve the issue you are having, then I would be happy to either send a replacement antibody or to process a refund.

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

Thank you for your email. customer is happy about the credit note offered.

ANSWER:

Thank you for confirming this information and for your help and cooperation with this case. As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. The reference number, or credit ID, is xxx. Please refer to this number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.