Anti-Integrin beta 1 antibody [EP1041Y] - Carboxyterminal end (ab52971)

Product code: 52971
Lot number: GR3780-1
Inquiry: Multi-bands These data were shown multi bands. so I would like to know what size is integrin beta 1. Please advice or comment about these data.
1) Abcam product code
ab52971
2) Abcam order reference number or product batch number
GR3780-1
2-1) storage temperature of antibody
-20°C
3) Description of the problem
Multi-bands
4) Sample preparation:
Type of sample and species(whole cell lysates, fraction, recombinant protein…and Human,
mouse…): Stem cell from huma
Lysis buffer
RIPA buffer
Protease inhibitors: Protease inhibitor cocktail
Phosphatase inhibitors: Yes
Reducing agent: DTT
Boiling for ≥5 min? yes
Protein loaded ug/lane or cells/lane 25ug
Positive control : no
Negative control : no
5) Percentage 8% gel
Type of membrane
: PVDF membrane
Protein transfer verified 90V 1hr 140V 1hr , Ponceau S
Blocking agent and concentration
5% skim milk
Blocking time
1hr
Blocking temperature
RT
6) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution 1:1000
Diluent buffer PBST or 5% skim milk
Incubation time overnight
Incubation temperature: 4 °C
7) Secondary antibody: (anti-rabbit IgG abcam ) no. SA002-500
Species:goat
Isotype: IgG
Reacts against: rasbbit
Concentration or dilution 1:5000
Diluent buffer 5% skim milk
Incubation time 1hr
Incubation temperature: RT
Fluorochrome or enzyme conjugate: HRP
8) Washing after primary and secondary antibodies:
Buffer 1X PBST
Number of washes: 5time 10min
9) Detection method
ECL solution
10) How many times have you run this staining? 2times
-Do you obtain the same results every time? yes
-Have you run a "No Primary" control? yes
-What steps have you altered to try and optimize the use of this antibody?
Diluent buffer of primary antibody

ANSWER:

Thank you for your enquiry regarding ab52971 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this antibody.

Q1: Could you please explain what the 1st and 2nd sets of experiments represent on the attached image?

Q2: It would be very useful for me to know what the different lanes (at least 3) demonstrate on the WB images.

Q3: Could you please clarify the stem cell type used as samples?

Q4: What positive control was applied in these experiments?

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

I safely received the ab5506 antibody and tried it out this weekend. I used 5% goat serum as a blocking agent this time. Unfortunately, it still produced no staining in either normal or tumour prostate. I have also tried Lrgi1 and Cxcr4 antibodies (ab2074 & ab36707) without antigen retrieval. To my surprise the staining worked, but it was still nuclear instead of predicted membrane localisation. So no improvement over the protocol with antigen retrieval. I think we'll have to go with the refund option.

ANSWER:

Thank you for you reply.

Your credit note IDs are ***** & *********.

I am sorry that these antibodies did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you for all of these antibodies (ab2074, ab5506, ab32392, ab39707, ab51317, ab52971 and ab65006).

The credit notes may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

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

We've purchased a number of antibodies from AbCam over the course of the past 6 months and we're struggling to get them to work properly. We therefore wonder if you could provide us with some advice to improve their performance. The antibodies in question and their respective dilutions are:
CXCR4 - ab2074 (1:100)
EpCam - ab32392 (1:250)
LRIG1 - ab36707 (1:200)
Integrin beta1 - ab52971 (1:250)
TACD2 - ab65006 (1:200)
c-Kit - ab5506 (1:100)
Sca1 - ab51317 (1:100)
Our samples are murine tumour and normal tissue from Skin, Mammary and Prostate (only Prostate for the last three antibodies). All samples are Formalin fixed and Paraffin embedded. We use a generic immunohistochemistry protocol. Antigen retrieval is performed by heating slides in Dako Citrate buffer at 100C for 20 min. TBS+Tween20 is used as washing solution. The blocking agent (for rabbit antibodies) is 10% normal goat serum. Antibody binding is detected by anti-rabbit HRP-conjugated secondary antibody from Envision kit with subsequent visualisation with DAB reagent. Rabbit anti-rat biotinylated secondary in 10% normal rabbit serum was used to detect Sca1 primary followed by ABC kit (vectastatin) and DAB treatment.

The main problems we've encountered with the named antibodies are the lack of positive staining (for c-Kit and Sca1 antibodies) and non-specific staining (for the rest). CXCR4, LRIG1, EpCam, Integrin beta 1 and TACD2 all displayed nuclear staining of various intensity instead of expected membrane-associated staining pattern.

We would therefore appreciate any suggestions on potential ways to improve the staining. Please, let me know if you would like me to send you detailed protocols and/or images of the stained tissue.

ANSWER:

Thank you for contacting Abcam.

I am sorry about all of the problems you have been having with these antibodies.

It would greatly be appreciated if you could send me some of the images that you are getting using these antibodies and also could you answer the questions below, so that I can understand your protocol a little more:

1 - How long do you block your samples for?

2 - How long do you incubate your primary antibodies for?

3 - Do you use a hydrogen peroxidase step?

4 - Are your samples permeabilised? If so with what percentage of detergent and for how long?

5 - Have you tried testing any of these antibodies without antigen retrieval, as for ab2074 & ab36707 at least, we have some information that suggests antigen retrieval is not necessary.

Once again, I am sorry about all the problems you have been and having and I hope to able to help you resolve this issue.

I look forward to your reply.

The latest lot of this product is not working.

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 .

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.

Dear technical team, Our customer would like to purchase 11 antibodies from abcam and needs control for each of antibody. The customer will test IHC-P so, control should be slide type. I have searched only one of them as below and would like to ask check the rest of them.  

ANSWER:

Thank you very much for contacting us - and sending the enquiry as a table. That clear overview was very helpful :-)

As you may know, we have not tested the individual samples for their expression of the mentioned targets. However, according to the literature and other sources such as http://www.proteinatlas.org/ these tissues exhibit high expression of these proteins and thus are suitable controls.

Cat# Product Positive control Slide ab27526 Anti-Notch1 antibody Breast carcinoma ab4697, ab4698 (but also ab4701, ab4700, ab4696) ab7970 Anti-NFkB p65 antibody Spleen (or skin) ab4372 (or ab4365, ab4417)　 ab10800 Anti-Integrin alpha 2 antibody Bladder, gall bladder, kidney　 ab4287, ab4330, ab4347　 ab20142 Anti-Integrin alpha 6 antibody Bladder, gall bladder, oesophagus　 ab4287, ab4330, ab4404　 ab16821 Anti-Integrin alpha V antibody Human breast carcinoma ab4697, ab4698 (but also ab4701, ab4700, ab4696)　 ab52971 Anti-Integrin beta 1 antibody Human breast carcinoma ab4697, ab4698 (but also ab4701, ab4700, ab4696)　 ab75872 Anti-Integrin beta 3 antibody Human spleen lysate and human tonsil tissue. ab4372, ab5146 ab79007 Anti-Integrin alpha 1 + beta 1 antibody Rat gut, heart and lung tissue. ab4620, ab4617　 ab30483 Anti-Integrin alpha 2+beta 1 antibody Bladder, gall bladder,breast carcinoma ab4287, ab4330, ab4697, ab4698 (but also ab4701, ab4700, ab4696)　 ab16822 Anti-Integrin alpha 2b + beta 3 antibody Spleen, kidney　 ab4372, ab4347　 ab78289 Anti-Integrin alpha V beta 3 antibody Bladder, oesophagus　 ab4287, ab4404　

All the above mentioned antibodies are guaranteed to work in immunohistochemistry on paraffin-embedded sections as stated on the datasheet. However, we do not currently have an image of ab10800, ab16821, ab30483, ab16822, and ab78289 in IHC-P. Therefore, I can offer a discount off a future purchase if you buy one of these antibodies now, use it IHC-P and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free primary antibody each. If you decide to go ahead and purchase this product, please contact our Scientific Support team prior to the purchase for more details.

I hope this information is helpful to you. Please do not hesitate to contact me if you need any further advice or information in this regard.