Overview

  • Product name
    Anti-Ctip1/BCL-11A antibody [14B5]
    See all Ctip1/BCL-11A primary antibodies
  • Description
    Mouse monoclonal [14B5] to Ctip1/BCL-11A
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, ICC, IP, IHC (PFA fixed), ChIP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
  • Immunogen

    Fusion protein corresponding to Ctip1/BCL-11A.

  • Epitope
    Epitope is in core of CTIP1/Bcl11a (aa's 172-434).
  • General notes

    Previously labelled as Ctip1.

Properties

Applications

Our Abpromise guarantee covers the use of ab19487 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 at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 91 kDa).
ICC Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC (PFA fixed) Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. PubMed: 23576758
Flow Cyt Use 0.1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Functions as a myeloid and B-cell proto-oncogene. May play important roles in leukemogenesis and hematopoiesis. An essential factor in lymphopoiesis, is required for B-cell formation in fetal liver. May function as a modulator of the transcriptional repression activity of ARP1.
  • Tissue specificity
    Expressed at high levels in brain, spleen thymus, bone marrow and testis. Expressed in CD34-positive myeloid precursor cells, B-cells, monocytes and megakaryocytes. Expression is tightly regulated during B-cell development.
  • Involvement in disease
    Note=Chromosomal aberrations involving BCL11A may be a cause of lymphoid malignancies. Translocation t(2;14)(p13;q32.3) causes BCL11A deregulation and amplification.
  • Sequence similarities
    Contains 6 C2H2-type zinc fingers.
  • Post-translational
    modifications
    Sumoylated by SUMO1.
  • Cellular localization
    Cytoplasm. Nucleus. Associates with the nuclear body. Colocalizes with SUMO1 and SENP2 in nuclear speckles.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2810047E18Rik antibody
    • B cell CLL/lymphoma 11A (zinc finger protein) antibody
    • B cell CLL/lymphoma 11A (zinc finger protein) isoform 2 antibody
    • B-cell CLL/lymphoma 11A antibody
    • B-cell lymphoma/leukemia 11A antibody
    • BC11A_HUMAN antibody
    • BCL-11A antibody
    • BCL11A antibody
    • BCL11A B cell CLL/lymphoma 11A (zinc finger protein) isoform 1 antibody
    • BCL11A L antibody
    • BCL11A S antibody
    • BCL11A XL antibody
    • BCL11a-M antibody
    • BCL11AL antibody
    • BCL11AS antibody
    • BCL11AXL antibody
    • C2H2 type zinc finger protein antibody
    • COUP TF interacting protein 1 antibody
    • COUP-TF-interacting protein 1 antibody
    • CTIP1 antibody
    • CTIP1, mouse, homolog of antibody
    • D930021L15Rik antibody
    • Ecotropic viral integration site 9 antibody
    • Ecotropic viral integration site 9 homolog antibody
    • Ecotropic viral integration site 9 protein antibody
    • Ecotropic viral integration site 9 protein homolog antibody
    • EVI-9 antibody
    • Evi9 antibody
    • Evi9, mouse, homolog of antibody
    • FLJ10173 antibody
    • FLJ34997 antibody
    • HBFQTL5 antibody
    • KIAA1809 antibody
    • mKIAA1809 antibody
    • OTTHUMP00000159788 antibody
    • OTTHUMP00000159789 antibody
    • OTTHUMP00000201250 antibody
    • OTTHUMP00000202084 antibody
    • Zinc finger protein 856 antibody
    • ZNF856 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Empty (0 µg)
    Lane 3: BCL11A (Ctip1) knockout HAP1 whole cell lysate (20 µg)
    Lane 4: Raji whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab19487 observed at 91 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab19487 was shown to specifically react with BCL11A (Ctip1) in HAP1 wild type cells. No band was observed when BCL11A (Ctip1) knockout cells were examined. Wild-type and BCL11A (Ctip1) knockout samples were subjected to SDS-PAGE. ab19487 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Ctip1/BCL-11A antibody [14B5] (ab19487)

    Lane 1 : C6 Cell Line
    Lane 2 : MEF Cell Line
    Lane 3 : HDFn Cell LIne
    Lane 4 : Jurkat Cell Line
    Lane 5 : HEK293T Cell Line

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 91 kDa



    The CTIP1 / BCL-11A protein band is indicated at 92 kDa. This antibody also identified a 55 kDa protein band in Western blotting, principally in mouse fibroblasts (MEF), but this is unrelated to.

     

    Blocking was performed in 5% milk (in PBS). The primary antibody was diluted in 1% milk (in PBS) and incubated overnight at 4ºC.

  • Overlay histogram showing Ramos cells stained with ab19487 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab19487, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Anti-Ctip1/BCL-11A antibody [14B5] (ab19487) at 1 µg/ml + Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 91 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa, 42 kDa, 62 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    The 120 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Ctip1 / BCL-11A.

References

This product has been referenced in:
  • Stankova T  et al. SUMO1-conjugation is altered during normal aging but not by increased amyloid burden. Aging Cell N/A:e12760 (2018). Read more (PubMed: 29633471) »
  • Shen S  et al. Predictors of lymphovascular invasion identified from pathological factors in Chinese patients with breast cancer. Oncotarget 9:2468-2474 (2018). Read more (PubMed: 29416785) »
See all 23 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

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

Answer

Thank you for your message.

I am sorry to hear the replacement vial has also not worked for you.I suggest it would be appropriate to offer you a credit note, refund,or an alternative replacement in this case.

I appreciate the time you have spent on this in the laboratory and apologize for the inconvenience. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1129389.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your message and for kindlyproviding this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange afree of charge replacementor credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

Order Details
Antibody code:ab19487

Problem
Choose: Non-specific band in prostate BPH clinical sample and no band in positive control: MDA231 cell line

Lot number GR5236-3

General Information
Antibody storage conditions (temperature/reconstitution etc): aliquoted, kept at -20C


Description of the problem (high background, wrong band size, more bands, no band etc.)
Non-specific band in prostate BPH clinical sample (around 30 kDa) and no band in positive control: MDA231 cell line


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
cell lines: MDA231, MCF7, Pnt1a, Pnt2, FTC133, TPC1, DLD1, HTC116 and 1 clinical prostate sample

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
standard: lysis buffer with protease inhibitors

Amount of protein loaded
30

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)

reducing conditions, 7.5%


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
2h at 200mA in standard transfer buffer (TRIS GLYCINE with 20% methanol).

Blocking: 5% milk in TRIS

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
BCL11a, mouse, ab19487 in 5% BSA/TRIS, overnight incubation

Wash: TRIS/tween 2x10min

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Fisher, 32430, goat anti-mouse peroxidise conjugated, 1:200 for 1h at room temp.
Wash: TRIS/tween 2x10min


Detection method (ECL, ECLPlus etc.)
chemiluminescent HRP substrate

Positive and negative controls used (please specify)
Positive: breast ca cell line MDA231, negative: MCF7. Based on information from collaborators who are about to publish their data. They used the same clone antibody but from Santa Cruz. However Santa Cruz discontinued their product.


Optimization attempts (problem solving)
How many times have you tried the Western?

3 times: incubated 1Ab longer, exposed film longer, shorter washes

Have you run a No Primary control?
Yes No

No as I have clean image anyway

Do you obtain the same results every time?
Yes No

More or less. When I re-incubated the membrane again overnight and exposed film for very long I got additional background bands but any at predicted size


e.g. are the background bands always in the same place?


What steps have you altered?
wash times, X ray exposure times

Additional Notes:
I have no problem with background. The image is clean but there is no specific band at predicted size at all.

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 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. I would also appreciate if you can confirm some further details:

1. I would appreciate if you are able to provide further details of the lysis buffer that was used? I can suggest to use RIPA if not already tried.

2. For loading, is this 30 micrograms?

3. Was the overnight incubation at4oC? This can often provide more efficient and specific staining if its not already been tried.

4. Has the protein transfer to the membrane and quality of the sample beenassessed using a loading control?

5. Unfortunately, I am sorry I am not able to open the image. I would appreciate if you could try sending this again.

6. Is the secondary antibody working well with other primary antibodies?

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 taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and human 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 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 a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if youare also able to provide an image, including molecular weight markers,which would help us to assess the results.

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. I can confirm that the immunogens for this antibody were GST-CTIP1 1-171, GST-CTIP1 172-434 and GST-CTIP1 420-776. The three immunogens were used in approximately equal amount to immunize the mice. ab19487 was then prepared from hybridomas that produced antibodies recognizing different regions of Ctip1. I can also confirm that ab19487 will bind bcl11a alone as well as any bcl11a (Ctip1) complex. I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

Read More

Answer

Thank you for your inquiry. All of the information we have for each of our products is listed on the online datasheet for your convenience. These are updated as soon as any new information is brought to our attention. I can confirm his monoclonal antibody was mapped to the region of core of CTIP1/Bcl11a (aa's 172-434). A closer mapping was not performed. This monoclonal antibody is produced by a hybridoma cell line and then purified to the IgG fraction. Should you decide to go ahead and purchase this product, we encourage you to submit an Abreview of your results. We appreciate all customer feedback, whether positive or negative, and we make all product information available to researchers. To find out more about our Abreview system, please see the following webpage: https://www.abcam.com/abreviews Please do not hesitate to contact us again if you need further assistance.

Read More
Application
ChIP
Sample
Mouse Cell lysate - nuclear (haematopoietic stem progenitor cell line)
Specification
haematopoietic stem progenitor cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 0.4% and 1% formaldehyde
Positive control
Gata1 promoter and b-globin HS3

Abcam user community

Verified customer

Submitted Oct 20 2010

Application
Immunohistochemistry (PFA fixed)
Sample
Mouse Tissue sections (Brain (P7 coronal section))
Specification
Brain (P7 coronal section)

Abcam user community

Verified customer

Submitted Feb 25 2008

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