Anti-Histone H3 antibody - ChIP Grade (ab12079)

Dear technical support team:

This customer has purchased ab1772 (Histone H3 peptide - di methyl K9) and has conducted the ELISA assay with ab12079 and ab32521. The result shows the OD value is too weak; therefore, he wants to ask for your help to modify his experiment step, could you please offer any suggestion to help him?

His experiment step as follow:



1) Abcam product code: ab1772



2) Abcam order reference number or product batch number Lot: GR34813-1

3) storage temperature -80℃

4) Description of the problem: the OD value is too weak

5) Type of ELISA: direct, indirect, sandwich sandwich

6) Sample details: ab1772

7) Target antigen (if found in heterogeneous sample) ab1772

8) Positive control Negative control

9) Blocking agent (eg BSA…): BSA

Concentration 1%

Blocking time 1 hour

Blocking temperature 25℃

10) Capture or Primary antibody information

Used to coat wells? Yes/no ab12079

Species Goat

Reacts against Hu, Ms

Concentration or dilution 1 μg/mL

Diluent buffer PBS

Incubation time overnight

Incubation temperature 25℃



11)Detection antibody: ab32521

Species Rabbit

Reacts against Hu, Ms

Concentration or dilution 1/10000

Diluent buffer 99% PBS, BSA

Incubation time 1 hour

Incubation temperature: 25℃

Conjugation ab1772



12) Enzyme labelled secondary antibody: ab6721

Species Goat

Reacts against Rabbit

Concentration or dilution 50 ng/mL

Diluent buffer 10 mg/mL BSA, 0.15 M Sodium chloride, 0.02 M Potassium phosphate

Incubation time 1 hour

Incubation temperature: 25℃

Conjugation ab113680



13) Washing after primary and secondary antibodies:Yes

Buffer 0.5% Tween 20, PBS

Number of washes 3 times

Length of washes 10 secs



14) Substrate Tetramethylbenzidine Substrate

Incubation time 30 mins



15) How many times have you run this ELISA? 2

Do you obtain the same results every time? yes

What steps have you altered to try and optimize the use of this antibody? No



Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

Ada




呂侑侖 (Ada Lu)
Technical Support
Interlab Co., Ltd (卓昇有限公司)
Phone: +886-2-27367100
Fax: +886-2-27359807

ANSWER:

Thank you for taking the time to complete our questionnaire and contact us. 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.

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

I would like to reassure you that 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.

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

1. I have checked whether these antibodies would detect the peptide:

ab1772 Histone H3 peptide - di methyl K9
Sequence: QTARK(Me2)STGG-C
I am sorry to confirm this peptide is not tested and covered by our guarantee in ELISA.

ab12079 Anti-Histone H3 antibody - ChIP Grade
Immunogen sequence: C-IQLARRIRGERA

I am sorry this antibody would not detect the ab1771 peptide, as its immunogen sequence does not contain the same amino acid sequence as the peptide. I can suggest this is why a signal is not being obtained. It has also not been tested in ELISA or sandwhich ELISA.

2. Could you confirm if any other samples have been tried and what the results were?

I am sorry we regrettably do not have a combination of antibodies that would be guaranteed to work to detect this peptide in sELISA. All tested and guaranteed applications are specified on our datasheets, and these are updated as soon as any new information is brought to our attention. Sandwich ELISA procedures can be difficult to optimize and tested match pair antibodies should be used. This ensures the antibodies are detecting different epitopes on the target so they do not interfere with the other antibody binding. Therefore, I regrettably we are unable to guarantee our antibodies in sandwich ELISA unless they have been specifically tested for sandwich ELISA.

I can suggest you may like to consider trying an indirect peptide ELISA with ab32521 or ab113680 which are more likely to work. Although I am sorry that as the peptide is not tested in ELISA, we would not be able to provide our guarantee for this.

I hope this information is helpful and helps to explain the results, thank you for your cooperation. Please do not hesitate to contact me again if you have any further questions or concerns regarding this case.

Dear technical support team:

This customer has purchased ab1771 (Histone H3 peptide - mono methyl K9) and has conducted the ELISA assay with ab12079 and ab113680. The results show no signal, therefore, he wants to ask for your help to modify his experiment step, could you please offer any suggestion to help him?

His experiment step as follow:

1) Abcam product code: ab1771

2) Abcam order reference number or product batch number Lot: GR71153-1


3) storage temperature -80℃



4) Description of the problem: no signal



5) Type of ELISA: direct, indirect, sandwich: sandwich



6) Sample details: ab1771



7) Target antigen (if found in heterogeneous sample): ab1771



8) Positive control Negative control





9) Blocking agent (eg BSA…): BSA

Concentration 1%

Blocking time 1 hour

Blocking temperature 25℃



10) Capture or Primary antibody information

Used to coat wells? Yes/no ab12079

Species Goat

Reacts against Hu, Ms

Concentration or dilution 1 μg/mL

Diluent buffer PBS

Incubation time overnight

Incubation temperature 25℃



11)Detection antibody:ab113680

Species Rabbit

Reacts against Hu, Ms

Concentration or dilution 100 ng/mL

Diluent buffer 99% PBS, BSA

Incubation time 1 hour

Incubation temperature: 25℃

Conjugation ab1771



12) Enzyme labelled secondary antibody: ab6721

Species Goat

Reacts against Rabbit

Concentration or dilution 50 ng/mL

Diluent buffer 10 mg/mL BSA, 0.15 M Sodium chloride, 0.02 M Potassium phosphate

Incubation time 1 hour

Incubation temperature: 25℃

Conjugation ab113680



13) Washing after primary and secondary antibodies:Yes

Buffer 0.5% Tween 20, PBS

Number of washes 3 times

Length of washes 10 secs



14) Substrate Tetramethylbenzidine Substrate

Incubation time 30 mins



15) How many times have you run this ELISA? 2

Do you obtain the same results every time? yes

What steps have you altered to try and optimize the use of this antibody? No



Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

Ada




呂侑侖 (Ada Lu)
Technical Support
Interlab Co., Ltd (卓昇有限公司)
Phone: +886-2-27367100
Fax: +886-2-27359807

ANSWER:

Thank you for taking the time to complete our questionnaire and contact us. 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.

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

I would like to reassure you that 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.

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

1. I have checked whether these antibodies would detect the peptide:

ab1771 Histone H3 peptide - mono methyl K9
Sequence: KQTARK(Me)STGG-C
I am sorry to confirm this peptide is not tested and covered by our guarantee in ELISA.

ab113680 Anti-Histone H3 (mono methyl K9) antibody?
Immunogen sequence: ARK(Me)ST
I can suggest this probably would detect the peptide. However, I am sorry to confirm this antiobdy is tested inindirect ELISA, andnot sandwhich ELISA and has not specifically been tested with this peptide.

ab12079 Anti-Histone H3 antibody - ChIP Grade
Immunogen sequence: C-IQLARRIRGERA
I am sorry this antibody would not detect the ab1771 peptide, as its immunogen sequence does not contain the same amino acid sequence as the peptide. I can suggest this is why a signal is not being obtained.

2. Could you confirm if any other samples have been tried and what the results were?

I am sorry we regrettably do not have a combination of antibodies that would be guaranteed to work to detect this peptide in sELISA. All tested and guaranteed applications are specified on our datasheets, and these are updated as soon as any new information is brought to our attention. Sandwich ELISA procedures can be difficult to optimize and tested match pair antibodies should be used. This ensures the antibodies are detecting different epitopes on the target so they do not interfere with the other antibody binding. Therefore, I regrettably we are unable to guarantee our antibodies in sandwich ELISA unless they have been specifically tested for sandwich ELISA.

I can suggest you may like to consider tryinganindirect peptide ELISA with ab113680 which is more likely to work. Although I am sorry that as the peptide is not tested in ELISA, we would not be able to provide our guarantee for this.

I hope this information is helpful and helps to explain the results, thank you for your cooperation. Please do not hesitate to contact me again if you have any further questions or concerns regarding this case.

Dear abcam technical support team:
This customer already purchase ab8895, ab32521, ab32356, ab12079, and conducted WB with human sample, but now he would likes to use these antibodies and peptides to test in ELISA assay with human sample, would you please help this customer to create a discount code?

ANSWER:

Thank you for contacting us.

ab71998 and ab71999 have already beentested in ELISA.

For the other antibodies, I am sorry we cannot issue testing discount codes for antibodies that have already been purchased.A testing discountcode must be issued before the order is placed. However, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases.

To find out more about our Abreview system, please see the following link:

http://www.abcam.com/abreviews

I hope this information is neverthelesshelpful. Please do not hesitate to contact us again with any other questions.

looking for H3 antibody for WB and yeast

ANSWER:

Thank you for contacting us.

I have checked our catalog and found the following 4 antibodies for histone H3 that have been tested in WB and yeast (either Saccharomyces cerevisiae and/orSchizosaccharomyces pombe):

ab1791: http://www.abcam.com/Histone-H3-antibody-ChIP-Grade-ab1791.html
ab12079: http://www.abcam.com/Histone-H3-antibody-ChIP-Grade-ab12079.html
ab18521: http://www.abcam.com/Histone-H3-antibody-ab18521.html
ab46765: http://www.abcam.com/Histone-H3-antibody-ChIP-Grade-ab46765.html

We guarantee all our products for tested species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: http://www.abcam.com/abpromise.

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: http://www.abcam.com/collaborationdiscount.

If you are in need of any other product that you might require for your experiments (e.g. secondary antibodies, controls), please let me know as well.

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