Overview

  • Product name
  • Description
    Rabbit polyclonal to Gata6
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 500 to the C-terminus of Human Gata6.

    Read Abcam's proprietary immunogen policy (Peptide available as ab24410.)

  • Positive control
    • Human embryonic stem cells differentiated into mesendoderm. This antibody also gave a positive signal in D3 ES Cells Day 15 EB (Gata6 +ve) lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab22600 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 1/200.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). Abcam recommends using milk as the blocking agent.
ICC/IF 1/200. PubMed: 20739751

Target

Images

  • Anti-Gata6 antibody (ab22600) at 1 µg/ml + D3 ES Cells Day 15 EB (Gata6 +ve) at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 45 kDa
    Observed band size: 45 kDa
    Additional bands at: 73 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 2 minutes
  • Image courtesy of Human Protein Atlas

    ab22600 staining Gata6 in Human stomach, with strong staining within the glandular cells. Paraffin embedded human stomach was incubated with ab22600 (1/80 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab22600 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .

References

This product has been referenced in:
  • Ye B  et al. Klf4 glutamylation is required for cell reprogramming and early embryonic development in mice. Nat Commun 9:1261 (2018). Read more (PubMed: 29593216) »
  • Zhou A  et al. Nuclear GSK3ß promotes tumorigenesis by phosphorylating KDM1A and inducing its deubiquitylation by USP22. Nat Cell Biol 18:954-966 (2016). Read more (PubMed: 27501329) »
See all 15 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Bos taurus Cell (Blastocyst)
Permeabilization
Yes - 0.5% Triton X-100 in PBS
Specification
Blastocyst
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Paraformaldehyde

Dr. Marcelo Goissis

Verified customer

Submitted Sep 01 2018

Question

Hi Sir: I attached the questionaire to this E mail. Thank you and hope to hear from you soon. Regards   Order Details Antibody code: ab22600 Problem Choose: Non-specific bands Lot number:  GR13245 (1MG/ML) Purchase order number: 6026197 or preferably Abcam order number: 983467 General Information Antibody storage conditions (temperature/reconstitution etc) -20 degree centigrade Description of the problem (high background, wrong band size, more bands, no band etc.) Several bands seen, including one around 70 kDa. None at 45 kDa as expected. Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)   Mouse heart lysate (homogenized) in different amount and early stage embryoid body lysate (sonicated). Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) RIPA(+protease inhibitor tablet) What was the lysis procedure? Mouse heart lysate (homogenized) in different amount and early stage embryoid body lysate (sonicated). Have the samples been reduced and denatured with sample buffer containing SDS and mercaptoethanol by heating for 5 mintues at 95oC? No Have you tried without sonication? Heart sample was homogenised Amount of protein loaded 20- 160 ug. Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) 12% gel (Introvirgin) Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Buffer: in 2ml : 22.5g Glycine  6.06g Tris  400ml Methanol  2gSDS Wet transfer (300mA 200mV) for 6 hours Blocking with 5% milk powder in TBST Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) (1:150) for 16 hours at 4 degree centigrade Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Licor:926-32211 IRDye 800CW Goat anti-Rabbit IgG (H + L), 0.5mg   Is the secondary antibody working well with other primary antibodies? Yes Detection method (ECL, ECLPlus etc.) Licor-odyssey Positive and negative controls used (please specify) No Optimization attempts (problem solving) Do different dilution of protein How many times have you tried the Western? 50 Have you run a "No Primary" control? No What were the results of the no primary control? Do you obtain the same results every time? Yes No e.g. are the background bands always in the same place? What steps have you altered? Additional Notes: Because I used Gata-6 antibody from Santa Cruz before this one. I cannot get a good signal from that one so I bought this one. I am pretty sure about my skill of western blotting.

Read More
Answer

Thank you for taking the time to complete our questionnaire. 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. Reviewing this case, I would like to offer some suggestions to help optimize the results from ab22600:   1. I can suggest to reduce and denature the sample by heating to 95oC for 5 minutes with sample buffer containing SDS and mercaptoethanol. The bands observed may be polymers of the protein which will run as single protein when reduced and denatured.  Also to run on a reducing gel, to ensure the sample is in the correct conformation to migrate at the correct molecular weight and be detected by the antibody.  2.  What are the results of the no primary control? 3.  We recommend to load 20 - 30 ug of protein per lane of a gel to ensure it is not overloaded. In the event that a product is not functioning in the applications 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. I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.  

Read More

Answer

Thank you for contacting Abcam. From the information on the datasheet for ab22600, the antibody does recognize band ay ~45kDa and also a band about ~75kDa, therefore I am unsure what the other band that you are detecting could be. For the peptide ab24410, it is 17 amino acid fragment from within the C-terminal end. Please let me know is there is anything else I can help you with or if you continue to have issues with ab22600.  

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (aorta)
Loading amount
12 µg
Specification
aorta
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Oct 29 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (differentiated mouse EBs (day 14) D3 line)
Loading amount
20 µg
Specification
differentiated mouse EBs (day 14) D3 line
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Jun 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Intestine)
Specification
Intestine
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 09 2008

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

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