Overview

  • Product name
  • Description
    Rabbit polyclonal to SOCS3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide (

    QLPGPIREFLDQYDAPL

    ), corresponding to amino acids 209-225 of the human/ rat/ mouse SOCS 3.

  • Positive control
    • Cardiac myocytes and A431 whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab3693 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
IHC-P Use a concentration of 2 - 20 µg/ml.
WB Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IP Use 3-5µg for 107 cells.

Target

  • Function
    SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction. SOCS3 is involved in negative regulation of cytokines that signal through the JAK/STAT pathway. Inhibits cytokine signal transduction by binding to tyrosine kinase receptors including gp130, LIF, erythropoietin, insulin, IL12, GCSF and leptin receptors. Binding to JAK2 inhibits its kinase activity. Suppresses fetal liver erythropoiesis. Regulates onset and maintenance of allergic responses mediated by T-helper type 2 cells. Regulates IL-6 signaling in vivo (By similarity). Probable substrate recognition component of a SCF-like ECS (Elongin BC-CUL2/5-SOCS-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Seems to recognize IL6ST.
  • Tissue specificity
    Widely expressed with high expression in heart, placenta, skeletal muscle, peripheral blood leukocytes, fetal and adult lung, and fetal liver and kidney. Lower levels in thymus.
  • Pathway
    Protein modification; protein ubiquitination.
  • Involvement in disease
    Note=There is some evidence that SOCS3 may be a susceptibility gene for atopic dermatitis linked to 17q25. SOCS3 messenger RNA is significantly more highly expressed in skin from patients with atopic dermatitis than in skin from healthy controls. Furthermore, a genetic association between atopic dermatitis and a haplotype in the SOCS3 gene has been found in two independent groups of patients.
  • Sequence similarities
    Contains 1 SH2 domain.
    Contains 1 SOCS box domain.
  • Domain
    The ESS and SH2 domains are required for JAK phosphotyrosine binding. Further interaction with the KIR domain is necessary for signal and kinase inhibition.
    The SOCS box domain mediates the interaction with the Elongin BC complex, an adapter module in different E3 ubiquitin ligase complexes.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residues after stimulation by the cytokines, IL-2, EPO or IGF1.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATOD4 antibody
    • CIS 3 antibody
    • CIS-3 antibody
    • CIS3 antibody
    • Cish3 antibody
    • Cytokine induced SH2 protein 3 antibody
    • Cytokine-inducible SH2 protein 3 antibody
    • E2a Pbx1 target gene in fibroblasts 10 antibody
    • EF 10 antibody
    • MGC71791 antibody
    • SOCS 3 antibody
    • SOCS-3 antibody
    • Socs3 antibody
    • SOCS3_HUMAN antibody
    • SSI 3 antibody
    • SSI-3 antibody
    • SSI3 antibody
    • STAT induced STAT inhibitor 3 antibody
    • STAT-induced STAT inhibitor 3 antibody
    • Suppressor of cytokine signaling 3 antibody
    see all

Images

  • Localization of ab3693 as the precipitated red signal, with a hematoxylin purple nuclear counterstain in cardiac myocytes.
  • A431 whole cell lysate was probed with rabbit anti-SOCS-3 ab3693 at 1:200 dilution. A431 whole cell lysate was probed with rabbit anti-SOCS-3 (ab3693) at 1:200 dilution.
  • ab3693 (2µg/ml) staining SOCS3 in human jejunum using an automated system (DAKO Autostainer Plus). Using this protocol there is staining of the cells within the lamina propria.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:
  • Roncero AM  et al. Contribution of JAK2 mutations to T-cell lymphoblastic lymphoma development. Leukemia 30:94-103 (2016). WB ; Human . Read more (PubMed: 26216197) »
  • Akram KM  et al. Alveolar epithelial cells in idiopathic pulmonary fibrosis display upregulation of TRAIL, DR4 and DR5 expression with simultaneous preferential over-expression of pro-apoptotic marker p53. Int J Clin Exp Pathol 7:552-64 (2014). IHC-P ; Human . Read more (PubMed: 24551275) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 27 Abreviews or Q&A

Abreviews
Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (primary human macrophage)
Specification
primary human macrophage
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 19 2013

Answer


In general it would be ideal to have a positive control as perhaps SOCS3 is not expressed in A549 cells. We have another antibody go SOCS3, ab16030, where the antibody shows detection of a band in a number of common tumor cell types, Hela, Jurkat, A431 and MOLT4. If you or a colleague are culturing any one of these cell lines, I think it would be useful to do a diagnostic blot with lysate from one of these cell lines.
However, if a band is still not apparent, I would be happy to send you a replacement vial of any SOCS3 antibody.
I hope this is helpful. Please feel free to contact me with any further questions.

Read More
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (A549 lung carcinoma cells)
Loading amount
150000 cells
Specification
A549 lung carcinoma cells
Gel Running Conditions
Reduced Denaturing (15 % acryl amide)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Mr. Michel Crameri

Verified customer

Submitted Sep 25 2012

Answer

Thanks for your email.


It was a good idea to use the "EasyBlocker" reagent. It makes it quite clear that there is no SOCS3 being pulled down (IP'd) using ab3693, although the antibody does function well for western blotting alone as shown in the 'Input' lane.

I am happy to offer a replacementor credit. If you would like a replacement, please let me know which catalog number you need.

Thanks!

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Macrophages in liver , kidney & tonsil)
Specification
Macrophages in liver , kidney & tonsil
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No

Abcam user community

Verified customer

Submitted Aug 10 2012

Answer

Thanks so much for providing the additional details. I just have a few more follow up questions:

3. Based on the included WB image, it seems SOCS3 is detected from the input via WB but not the IP/WB. Do you agree? NO
==>If you could, please provide further details to support your reply.

5. How much cell lysate was incubated with the SOCS3 antibody during the IP?
3ug/ml
==>I understand that the IP antibody, ab3693, was added to the protein extract at 3ug antibody per ml of extract. Am I correct that the protein extract was also 3ug/ml in concentration? Usually protein extracts have a much higher protein concentration.

Thanks again for the additional information!

Read More

Question

- Abcam product code: ab3693
- Antibody storage conditions (temperature/reconstitution etc) -20℃
2. Please describe the problem (high background, wrong band size, more bands, no band etc). No signal
3. On what material are you testing the antibody in WB?
- Species:
- What’s cell line or tissue: homo oral cancer cell
- Cell extract or Nuclear extract: Cell extract - Purified protein or Recombinant protein: No 3.
The lysate - How much protein was loaded: 1.5 mg/ml
- What lysis buffer was used: RIPA buffer
- What protease inhibitors were used:
- What loading buffer was used:
- Phosphatase inhibitors Yes - Did you heat the samples: temperature and time: 4℃ O/N Matrix used to allow precipitation of antibody-antigen complex (Staphilococcus Aureus, Protein A Sepharose, Protein G, Protein A/G, or a second antibody) Protein G Detergent present in your system (Nonidet P40, SDS solution or other) SDS solution
4. Electrophoresis/Gel conditions/ Transfer conditions
- Reducing or non reducing gel: - Reducing agent:
- Gel percentage : 10% SDS-PAGE
- Transfer conditions: (Type of membrane, Protein transfer verified): PVDF , wet
5. Blocking conditions
- Buffer: 0.1% TBST
- Blocking agent: milk, BSA, serum, what percentage: 5% MILK
- Incubation time: 1 hr
- Incubation temperature: RT
6. Primary Antibody - Species: Rabbit - Reacts against:
- At what dilution(s) have you tested this antibody: 1:1000
- What dilution buffer was used: 5% BAS in 0.1% TBST
- Incubation time: O/N - Incubation temperature: 4℃ - What washing steps were done: 0.1% TBST 5 min 3 time -
7. Secondary Antibody - Species: - Reacts against:
- At what dilution(s) have you tested this antibody:
- Incubation time: 1 hr
- Wash steps: 0.1% TBST 5 min 3 time
- Fluorochrome or enzyme conjugate:
- Do you know whether the problems you are experiencing come from the secondary?
8. Detection method ECl, ECl+, other detection method: ECl
9. Did you apply positive and negative controls along with the samples? Please specify.
10. Optimization attempts - How many times have you tried the Western? 2 times
- Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no
- Do you obtain the same results every time e.g. are background bands always in the same place? yes
- What steps have you altered? Concentration of primary antibody

Read More
Answer

Thank you for your enquiry and for including a western blot image. I just have a few additional questions:

1. This seems to be an IP/WB experiment. Is this the case?

2. Is ab3693 used for both IP and WB? If not, which antibodies are used for IP and WB?

3. Based on the included WB image, it seems SOCS3 is detected from the input viaWBbut not the IP/WB. Do you agree?

4. SOCS3 band will run right around the 25kDa light and heavy chains from antibody. Its possible detection of the IP antibody light chain will obscure detection of SOCS3. What do you think abou this possibility?

5. How much cell lysate was incubated with the SOCS3 antibody during the IP?

6. How much antibody anti-SOCS3 antibody was incubated in the IP with the oral cancer cell extract?

7. What percentageSDS was present in the cell extract used ans input for IP?

Thanks in advance for this additional information.

Read More

Answer

Thank you for your reply.

I was wandering if the fellow who ordered the product would be able to provide some of the original order information? As I mentioned in my previous emails, I am willing to extend the 6 month Abpromise for a few months, I just need to get hold of the original order information. I do apologize that I cannot find it on my end, but if you can provide any information to help track it down, it would be very welcome.

I hope you are having a great day.

Read More

Question
Answer

Thank you for your reply.

I understand your situation, but I do need to have some indication about when ab3693 was actually ordered. A date, name or PO number would work.

As I mentioned previously, I am willing to extend the Abpromise for a few months in this situation, but I do need some information about the original order.

If you can provide me with any information regarding who placed the order, or the date it was placed, that would help me track down the order. I have now looked at all orders within the past year and I cannot find your one. I may be missing it and so any information you can provide will be helpful.

Once again, I look forward to your reply and helping resolve this issue.

Read More

1-10 of 27 Abreviews or Q&A

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

Sign up