Overview

  • Product name
    Anti-Guanylyl Cyclase alpha 1 antibody
    See all Guanylyl Cyclase alpha 1 primary antibodies
  • Description
    Rabbit polyclonal to Guanylyl Cyclase alpha 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Cow, Human
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide:

    KDAEDGNANFLGKASGVD

    conjugated to KLH by a Glutaraldehyde linker, corresponding to amino acids 673-690 of Rat Guanylyl Cyclase alpha 1

  • Positive control
    • Rat brain cytosolic fraction and trypsin digested, paraffin-embedded human, bovine and mouse heart sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab50358 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 1/10000. Detects a band of approximately 80 kDa (predicted molecular weight: 83 kDa).
IP Use at an assay dependent concentration.

5-10µg for 60-120µg of cytosolic fraction of rat brain.

IHC-P 1/100.

Target

  • Sequence similarities
    Belongs to the adenylyl cyclase class-4/guanylyl cyclase family.
    Contains 1 guanylate cyclase domain.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • GC S alpha 1 antibody
    • GC SA3 antibody
    • GCS alpha 1 antibody
    • GCS alpha 3 antibody
    • GCS-alpha-1 antibody
    • GCS-alpha-3 antibody
    • GCYA3_HUMAN antibody
    • Guanylate cyclase 1 soluble alpha 3 antibody
    • Guanylate cyclase soluble subunit alpha 1 antibody
    • Guanylate cyclase soluble subunit alpha 3 antibody
    • Guanylate cyclase soluble subunit alpha-3 antibody
    • Guanylate cyclase, nitric oxide-sensitive, alpha-1 subunit antibody
    • Guc1a1 antibody
    • GUC1A3 antibody
    • GUCA3 antibody
    • GUCSA3 antibody
    • Gucy1a1 antibody
    • Gucy1a3 antibody
    • MYMY6 antibody
    • nitric oxide-sensitive Guanylate cyclase, alpha-1 subunit antibody
    • NOGC, alpha-1 subunit antibody
    • Soluble guanylate cyclase large subunit antibody
    see all

References

This product has been referenced in:
  • Zhan R  et al. Nitric oxide induces epidermal stem cell de-adhesion by targeting integrin ß1 and Talin via the cGMP signalling pathway. Nitric Oxide 78:1-10 (2018). WB . Read more (PubMed: 29698689) »
  • Vogel S  et al. Platelet-derived HMGB1 is a critical mediator of thrombosis. J Clin Invest 125:4638-54 (2015). IP, ICC/IF ; Mouse, Human . Read more (PubMed: 26551681) »
See all 9 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Hela, Hep G2)
Specification
Hela, Hep G2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 05 2015

Question

Please find my completed questionnaire. I have also attached the powerpoint slide containing my western images.



I look forward to hearing from you.

Kind regards



1) Abcam product code: ab50358 and ab84955



2) Abcam order reference number or product batch number GR57221-2 and GR64869-2



3) Description of the problem: when running my various lysates for each of the antibodies listed above, the two westerns seem very similar in that the banding pattern seems the same, which triggers my concern over the specificity.



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…): tissue whole cell lysates

Lysis buffer: 62.5mMTris – HCl

2% SDS

10% Sucrose



Protease inhibitors: Sigma P8340

Phosphatase inhibitors: Sigma P5726

Reducing agent: beta mercaptoethanol

Boiling for ≥5 min? yes

Protein loaded ug/lane or cells/lane: 5ug for ab50358 and 10ug for ab84955

Positive control

Negative control



5) Percentage of gel: 10%

Type of membrane: methanol activated PVDF

Protein transfer verified: yes

Blocking agent and concentration: ab50358 5% non fat milk in TBS-T, ab84955 5% BSA in TBS-T

Blocking time: 1hr RT for ab50358 and overnight 4C for ab84955

Blocking temperature: as above



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution: ab50358 1:10,000 and ab84955 1:1000

Diluent buffer: ab50358 1% non fat milk, ab84955 1% BSA

Incubation time: ab50358 overnight, ab84955 1hr

Incubation temperature: ab50358 4C, ab84955 RT



7) Secondary antibody:

Species: Goat

Reacts against: Rabbit

Concentration or dilution: ab50358 1:5000, ab84955 1:1250

Diluent buffer : ab50358 1% non fat milk, ab84955 1% BSA

Incubation time: 1hr

Incubation temperature: RT

Fluorochrome or enzyme conjugate:



8) Washing after primary and secondary antibodies:

Buffer: TBS-T

Number of washes: 3 x 5 minutes



9) Detection method: Amersham ECL plus



10) How many times have you run this staining? 4

Do you obtain the same results every time? Yes

What steps have you altered to try and optimize the use of this antibody? Different primary antibody concentrations, different incubation times and temperatures, different block and diluents buffers, different secondary antibody concentrations



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.



Please send this questionnaire by e-mail by cutting and pasting the text in your e-mail (no attachments other than images) to technical@abcam.com.

Read More
Answer

Thank you for your enquiry regarding ab50358 and ab84955 and for taking the time to provide some useful details of the experiments.

I can confirm that these two antibodies are from two different sources and the immunogen sequences are also different.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

The protocol looks absolutely fine to me. The band patterns on the WB image look similar but not identical.

Lane 1: it looks like not enough material was loaded,

Lane 2 and 3: there may be protein degradation in the samples,

Lane 5: Major band for Guanylyl Cyclase alpha 1 at 70 kDa whilst for Guanylyl Cyclase beta 1 is at 60 kDa

Although, lysis buffer with proteinase inhibitors were used during sample preparation, protein degradation could still be an issue. In order to be able to prevent any enzymatic degradation, it is important to process the samples as quickly as possible after dissection. Is the human material is from biopsy samples or from postmortem tissues? Have you stored the tissues on ice before and during preparation?

Interestingly, for Guanylyl Cyclase beta 1 two different isoforms have been isolated and reported. You may wish to take a look at this site for further information:

http://www.uniprot.org/uniprot/Q02153

I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Answer

I am sorry to hear that you have been experiencing problems using these two products in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Please could you provide some further details of the protocol used and complete the following form (attached as a word document).

I am particularly interested in the followings:

- Sample preparation,

- Lysis buffer,

- Detection,

- No primary control - only secondary antibody

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

Read More

Question
Answer

DISCOUNT CODE: ***********
Expiration date: ************
I am very pleased to hear you would like to accept our offer and test ab50358 in IIHC-Fr. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-Fr 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: www.abcam.com/collaborationdiscount.

Read More

Question
Answer

Thank you very much for your interest in ab50358.
To our knowledge, ab50358 has not been tested in IHC-Fr. Therefore, I can offer a discount off a future purchase if you buy ab50358 now, test it in IHC-Fr and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free PRIMARY ANTIBODY.
If you are interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know that you would like to proceed and test ab50358 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab50358 so please wait for my reply before ordering.
2. Purchase ab50358 either by phone, fax, or online (www.abcam.com).
3. Test it in IHC-Fr.
4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.
5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.
We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab50358 turns out to be unsuitable for IHC-Fr, you will still receive the discount on your next purchase after your Abreview has been submitted.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Sheep Tissue lysate - whole (Sheep lung tissue)
Loading amount
40 µg
Specification
Sheep lung tissue
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Oct 01 2007

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

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