Overview

  • Product name

  • Description

    Rabbit polyclonal to DBX1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse
    Predicted to work with: Rat, Cow, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse DBX1 aa 100-200 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab103270)

  • Positive control

    • This antibody gave a positive signal in the following mouse tissue lysates: E14 Embryo Brain; E14 Embryo Spinal Cord; E16 Embryo Brain; E16 Embryo Spinal Cord; E18 Embryo Brain; E18 Embryo Spinal Cord. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: NIH-3T3.

Properties

Applications

Our Abpromise guarantee covers the use of ab61488 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 36 kDa).

Target

  • Relevance

    Dbx1 homeodomain transcription factor is expressed in progenitors at the boundary between the dorsal and ventral plates of the caudal neural tube, from which postmitotic cells migrate tangentially to their final destination. Dbx1 is implicated in patterning the central nervous system during embryogenesis. Cell fate allocation and cell diversity are determined at very early stages in progenitor cells at precise coordinates along the dorsoventral and anteroposterior axis. In the spinal cord, the spatially restricted expression of Dbx1 in progenitors is critical in establishing interneuron cell fates and helps coordinate diverse phenotypic features. In the telencephalon, Dbx1 is expressed in restricted progenitor domains at the borders of the developing pallium.
  • Cellular localization

    Nuclear
  • Database links

  • Alternative names

    • dbx antibody
    • Developing brain homeobox 1 antibody
    • Developing brain homeobox protein 1 antibody
    • Homeobox protein DBX1 antibody

Images

  • All lanes : Anti-DBX1 antibody (ab61488) at 1 µg/ml

    Lane 1 : E14 Mouse Embryo Brain Tissue Lysate
    Lane 2 : E14 Ms Embryo Spinal Cord Tissue Lysate
    Lane 3 : E16 Ms Embryo Brain Tissue Lysate
    Lane 4 : E16 Ms Embryo Spinal Cord Tissue Lysate
    Lane 5 : E18 Ms Embryo Brain Tissue Lysate
    Lane 6 : E18 Ms Embryo Spinal Cord Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 39 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 73 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • ICC/IF image of ab61488 stained NIH-3T3 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab61488 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

ab61488 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 22 2014

Question
Answer

Thank you for your reply.

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

Credit note ID: ######

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

Question

Dear Ms Hayes,
Please see my answers within the questionnaire below.
Kind regards,

Order Details
Antibody code: ab61488
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number GR35985-1

General Information
Antibody storage conditions (temperature/reconstitution etc) antibody aliquoted and stored at -20 according to manufacturer's recommendation upon arrival.

Description of the problem (high background, wrong band size, more bands, no band etc.) Non-specific band Multiple bands No signal or weak signal High background

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Total protein extract from mouse embryonic stem cells differentiated towards motor neuron (embryonic bodies). Testing for endogenous protein. We have confirmed that the gene is expressed in these cells by RNA-seq and RT-qPCR, repeated several times.

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Normal cell lysis buffer with medium stringency (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 1% Triton X-100) complemented with protease and phosphatase inhibitors from Roche. Samples mixed with sample buffer (60 mM Tris-Cl pH 6.8, 2% SDS, 10% glycerol, 5% β-mercaptoethanol, 0.01% bromophenol blue), heated at 70 for 10 min.

Amount of protein loaded 20 ug of sample loaded

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) NuPAGE Novex 4-12% Bis-Tris Gels under denaturing conditions, 180V for 1.5hrs

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Tris-glycine buffer for 2-3 hrs at 65V, blocked with 5% BSA first time and 5% milk second time.

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Dbx1 from Abcam rabbit, diluted 1:1000 first time and 1:500 second time in 5% BSA with 1% and 2% normal goat serum o/n at 4. Washed 3 x 10 min with PBS-Tween 0.5%

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Calbiochem Goat anti rabbit diluted 1:5000 in 5% BSA incubated 1 hr at RT followed by 3x5 min wash in PBS-T and 1x5 min in PBS

Detection method (ECL, ECLPlus etc.) ECL

Positive and negative controls used (please specify) As mentioned above, samples have been tested by RNA-seq

Optimization attempts (problem solving)

How many times have you tried the Western? 2 times

Have you run a No Primary control?
No, but we have not had with this secondary which we have been using for the past 3 years

Do you obtain the same results every time?
Yes
e.g. are the background bands always in the same place? Yes

What steps have you altered? Blocking, ab concentration
Additional Notes:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More
Answer

Thank you for taking the time to complete our questionnaire. The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note 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

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, ICC-F and for mouse samples. In the event that a product is not functioning in the tested applications and species 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 can confirm that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed technical questionnaire below. To keep it less time consuming for now, I have just included a WB questionaire. I would appreciate if you could complete this. It will help you put the information we require together very easily.
Thank you for your time and cooperation. We look forward to receiving the completed questionaire.
Order Details
Antibody code:
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Yes No
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:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Answer

Thank you for your reply. Frozen sections are falling in our category of immunohistochemistry (IHC-Fr). Please keep this in mind when submitting your Abreview.
Mouse samples have been tested, but human, rat and cow samples are only predicted to react (thus fall under the category "untested species").


DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab61488 in IHC-Fr. This code will give you 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for IHC-Frand 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: https://www.abcam.com/collaborationdiscount.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Zebra finch Tissue sections (embryonal spinal cord)
Specification
embryonal spinal cord
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TX100 in PBS
Blocking step
Roti-Immunoblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Dr. Robert Storm

Verified customer

Submitted Feb 06 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up