Overview

  • Product name
    Anti-Caveolin-1 (phospho Y14) antibody
    See all Caveolin-1 primary antibodies
  • Description
    Rabbit polyclonal to Caveolin-1 (phospho Y14)
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for Caveolin-1 only when phosphorylated at tyrosine 14.
  • Tested applications
    Suitable for: WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Cow, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Human Caveolin-1.
    Database link: Q03135

  • Positive control
    • NIH 3T3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab38468 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
WB 1/500 - 1/1000. Detects a band of approximately 20 kDa (predicted molecular weight: 20 kDa).
ELISA 1/20000.
ICC/IF 1/250.

Target

  • Function
    May act as a scaffolding protein within caveolar membranes. Interacts directly with G-protein alpha subunits and can functionally regulate their activity (By similarity). Involved in the costimulatory signal essential for T-cell receptor (TCR)-mediated T-cell activation. Its binding to DPP4 induces T-cell proliferation and NF-kappa-B activation in a T-cell receptor/CD3-dependent manner. Recruits CTNNB1 to caveolar membranes and may regulate CTNNB1-mediated signaling through the Wnt pathway.
  • Tissue specificity
    Expressed in muscle and lung, less so in liver, brain and kidney.
  • Involvement in disease
    Defects in CAV1 are the cause of congenital generalized lipodystrophy type 3 (CGL3) [MIM:612526]; also called Berardinelli-Seip congenital lipodystrophy type 3 (BSCL3). Congenital generalized lipodystrophies are autosomal recessive disorders characterized by a near absence of adipose tissue, extreme insulin resistance, hypertriglyceridemia, hepatic steatosis and early onset of diabetes.
  • Sequence similarities
    Belongs to the caveolin family.
  • Post-translational
    modifications
    The initiator methionine for isoform Beta is removed during or just after translation. The new N-terminal amino acid is then N-acetylated.
  • Cellular localization
    Golgi apparatus membrane. Cell membrane. Membrane > caveola. Membrane raft. Colocalized with DPP4 in membrane rafts. Potential hairpin-like structure in the membrane. Membrane protein of caveolae.
  • Information by UniProt
  • Database links
  • Alternative names
    • BSCL3 antibody
    • CAV antibody
    • CAV1 antibody
    • CAV1_HUMAN antibody
    • caveolae protein, 22 kD antibody
    • caveolin 1 alpha isoform antibody
    • caveolin 1 beta isoform antibody
    • Caveolin 1 caveolae protein 22kDa antibody
    • Caveolin-1 antibody
    • Caveolin1 antibody
    • cell growth-inhibiting protein 32 antibody
    • CGL3 antibody
    • LCCNS antibody
    • MSTP085 antibody
    • OTTHUMP00000025031 antibody
    • PPH3 antibody
    • VIP 21 antibody
    • VIP21 antibody
    see all

Images

  • Lanes 1-2 : Caveolin-1 Antibody
    Lanes 3-4 : Anti-Caveolin-1 (phospho Y14) antibody (ab38468) at 1/500 dilution

    Lane 1 : NIH 3T3 cell extract
    Lane 2 : NIH 3T3 cell extract. Pre-incubated with synthesized peptide
    Lane 3 : NIH 3T3 cell extract.
    Lane 4 : NIH 3T3 cell extract. Treated with H2O2.

    Predicted band size: 20 kDa
    Observed band size: 20 kDa

References

This product has been referenced in:
  • Bracamontes CG  et al. The serum protein profile of early parity which induces protection against breast cancer. Oncotarget 7:82538-82553 (2016). WB ; Human . Read more (PubMed: 27769065) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for your response.

At this stage it is difficult to identify the source of the problem and to be sure why this antibody failed to work in Western blot application.

It would be interesting to know 1) how well the vial was centrifuged before preparing the aliquots 2) whether the customer has used exactly the same aliquot for IHC and WB (head-to-head) or 3) separate vials for these two applications.

In future, I would like to advice and encourage the customer to aliquot the product in no smaller than 10 µl; the smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial.

The other thing which may be important to consider is the blocking agent. As I understand from the previous e-mail, the blots were blocked for 1h with 5% skim milk and then incubated with the primary antibody (1:1000 – as from the tested conditions) overnight at 4 degrees. Is that correct? How were the slides blocked for IHC/ICC?

Was milk applied as blocking agent for IHC/ICC as well or BSA or different solution?

Normally, for detection of phospho protein we would suggest using 5% BSA solution - as blocking - rather than milk. Milk may lead to significant decrease or loss of signal intensity due to reaction of primary antibody with blocking/diluting agent.

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

Read More

Answer

Thank you for your response.

My colleague is out of office for the rest of the week. I have tried to review this case and noticed that 2 ul aliquots were prepared. This may contribute to the problem your customer is facing with.

The size of the aliquots will depend on how much one typically uses in an experiment. Aliquots should be no smaller than 10 µl; the smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial.

I would advise to read this guide carefully, it has useful information storage.

https://www.abcam.com/index.html?pageconfig=resource&rid=10795

If you need anything further or any help then please let me know.

Read More

Question

This email has 2 sections:
A recap of the complaint
The respondent’s survey results (section-by-section)
Date(s) submitted: April 27, 2012
Catalog number(s): 38468
Product(s): Rabbit polyclonal to Caveolin 1 (phospho Y14)
CCEID: 3721273
Survey results
Telephone section
Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
A. No
Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
A.
Protocol advice section
Q. Did Abcam's Scientific Support give you protocol advice?
A.Yes
Q. How did you feel about being given protocol advice?
A.I was not expecting to be given protocol advice, but was happy to receive this
Q. Did the product work successfully after following the advice?
A.No
Q. Why have you not tried the protocol advice? (Select all that apply.)
A.
Free of charge replacement section
Q. Did Abcam's Scientific Support give you a free of charge replacement?
A. No
Q. Did the replacement product work successfully?
A.
Credit note / refund section
Q. Did Abcam's Scientific Support give you a credit note or a refund?
A.Yes
Q. Would you have preferred a different outcome from receiving a credit note or refund?
A.refund
Overall satisfaction section
Q. How satisfied are you with how the complaint was handled?
A.Neither satisfied nor unsatisfied
Q. Could we have done anything to better resolve the problem?
A.better antibody????
Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
A.No, I would not mind
Q. Please provide a phone number or email address where we can reach you.
Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.

Read More
Answer

Thank you very for participating in Abcam complaint survey.

We have provided the credit noteof full price; you can buy another antibody with that credit note without paying anything. I hope you have done that.

We had alsotried to solve the case however we were not able to determine the reason, why this antibody was not detecting band in WB as it should though it was OK in ICC/IF.

So do you need any further assistance? If yes, please let us know, we will be happyto help.

Read More

Answer

Thank you for contacting us.

Your credit note ID is XXXXXX.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note 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

Read More

Answer

Thank you very much for your cooperation. I am sorry to hear this antibody is not providing satisfactory results.

Having reviewed the protocol details again, I believe this product should have given satisfactory results. It is regrettable the results have not been successful.

I apologize for the inconvenience and am pleased to offer you a credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (uterus)
Specification
uterus
Fixative
no fixation
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 07 2012

Answer

Thanks you for your email.

This antibody should bind to reduced or denatured form, because we got positive binding for Western blots with the lysates of HuvEc and Hela cells. We haven’t received complains about this antibody. We didn’t use the antibody for tissue lysates. The phosphorylation on this site may be low in liver and ovary. So we recomend trying HuvEc lysates as positive control.

I hope this information will be helpful. HSould you have any other question please do not hesitate to contact us.

Read More

Question

Please see below for a customer’s technical enquiry
WESTERN BLOT
ANTIBODY DETAILS
Antibody Code: AB38468
Batch Number: GR13222
Order number: PO-11582
Abcam invoice number: 1285358
GENERAL INFORMATION
Antibody Storage Conditions (temperature/reconstitution etc)
Stored at -20 degrees : 2 uL aliquots to avoid repeat freeze thaw cycles
Description of the Problem (high background, wrong band size, more bands, no bands etc)
It works fine for immunofluorescence on cryosections – a technique that had not been tested.
The antibody from the spec details had been tested for western blotting. It however does not work at all for this technique. There are no bands present at all.
Sample (Species/Cell extract/Purified protein/Recombinant protein, etc)
Rat uterus – tissue lysates
Sample Preparation (Buffer/Protease inhibitors/Heating sample etc.)
Tissue is collected and immediately homogenized with lysis buffer (SDS, DOC, Igepal, NaCl, EDTA), aliquoted and protein estimated and stored at -80degrees until used for western blotting.
Samples are prepared with a loading buffer containing b-mercapto ethanol and boiled for 5minutes prior to loading on a gel.
Samples are then run at 200 V for 45 min; and transferred to PVDF membrane for 1h at 100V.
Blots are blocked for 1h with 5% skim milk and then incubated with the primary antibody (1:1000 – as from the tested conditions) overnight at 4 degrees
Blot is washed 3 times with 1x TBST
Blot is incubated with secondary antibody (1:2000 dil) – goat anti rabbit HRP for 2h at room temperature
Blot is washed 3 times with 1 x TBST
Blot is then imaged with ECL+ with the chemidoc imager from BioRad
Troubleshooting steps from the procedure set above include:
Loading more protein – 20ug – 60 ug
Using different loading conditions:
1) B mercapto ethanol + heat
2) B mercapto ethanol no heat
3) No b mercapto ethanol + heat
4) No B mercaptoethanol no heat
Tried using a different blocking percentage of milk 0.5% - 2 % and even not blocking at all.
Amount of protein loaded
20 – 60ug of protein
Electrophoresis/Gel Conditions (Reducing or non-reducing gel, % of the gel etc.)
Have tried using reducing and non reducing conditions – percentage of the gel has been kept the same – 12%
Transfer and blocking conditions (Buffer/time period, Blocking agent etc)
Blocking conditions: 0.5% - 5% skim milk 1h at room temperature – have tried even not blocking membrane at all
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
Abcam p-cav1 (Y14)/ rabbit/ 1X TBST, 0.5% skim milk/ 1:1000 (according to tested conditions – have tried using a 1:250 dilution)/ incubation time has been overnight at 4 degrees
Wash 3x with 1X TBST
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
Dako/ goat ant rabbit HRP/ 0.5 % skim milk/ 1:2000/2 hours at room temperature/ wash 3 x with 1X TBST
Detection Method (ECL, ECL Plus etc.)
ECL plus
Positive and negative controls used (please specify)
Positive controls have been kidney. – antibody works perfectly fine in immunofluorescence for the tissue being studied.
OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
How many times have you tried the Western Blot?
More than I can count
Have you run a “No Primary” control? No (Delete one)
Do you obtain the same results every time? Yes (Delete one)
: No bands are present
What steps have you altered?
I have as outlined above – altered the dilution range drastically to a 1:250 dilution.
I have tried lysing my tissue with a lysis buffer with no SDS or DOC with the same conditions as has been outlined.

Read More
Answer

Thank you for your email. I am sorry to hear that you are experiencing problems with this antibody.

I have read the protocol which looks fine to me however I would like to suggest few changes which will help to understand the cause of the problem;

This antibody has not been tested with rat lysates so we currently do not hold any rat and human comparison data. Although rat species is a predicted species however this is totally based on assumption of sequence homology only. Our recommendations is to try a human (HUVEC cells) or mouse kidney cell lysates treated with PMA 125ng/ml for 30 minutes as positive control.

The ab38468 antibody is a phospho specific antibody you may need to induce the phosphorylation of the Caveolin protein.

In uterus this protein is expressed at low level so we recommend using lysates of liver, ovary (stroma cells), HUVEC cells, Hela cells (PMA treated) as positive control.

I hope these suggestion will help to improve results. Should you have any other question please do not hesitate to contact us.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Cow Cell lysate - whole cell (Bovine Brain microvascular endothelial cells)
Loading amount
40 µg
Specification
Bovine Brain microvascular endothelial cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Cyril Mougin

Verified customer

Submitted Feb 11 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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