Overview

  • Product name

    Anti-MEIS1 antibody - ChIP Grade
    See all MEIS1 primary antibodies
  • Description

    Rabbit polyclonal to MEIS1 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, CHIPseq, ChIP, WBmore details
  • Species reactivity

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

    Synthetic peptide corresponding to Human MEIS1 aa 200-300 (internal sequence) conjugated to keyhole limpet haemocyanin.

  • Positive control

    • This antibody gave a positive signal in the following nuclear cell extract: KG-1. This antibody gave a positive signal in the following FFPE tissue: Human tonsil, and Rat normal brain.
  • General notes

    For a detailed method for MEIS1 ChIP-seq in RA-treated P19.6 cells, please see the protocol described by Dr Aurelien Serandour at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM819083.

Properties

Applications

Our Abpromise guarantee covers the use of ab19867 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 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
CHIPseq Use at an assay dependent concentration. PubMed: 22730288
ChIP Use at an assay dependent concentration. PubMed: 20971928
WB Use a concentration of 1 µg/ml. Detects a band of approximately 40, 48 kDa (predicted molecular weight: 43, 50 kDa). Abcam recommends using milk as the blocking agent.

Target

  • Function

    Acts as a transcriptional regulator of PAX6. Acts as a transcriptional activator of PF4 in complex with PBX1 or PBX2. Required for hematopoiesis, megakaryocyte lineage development and vascular patterning. May function as a cofactor for HOXA7 and HOXA9 in the induction of myeloid leukemias.
  • Tissue specificity

    Expressed at low level in normal immunohepatopoietic tissues, including the fetal liver. Expressed in a subset of myeloid leukemia cell lines, with the highest expression seen in those with a megakaryocytic-erythroid phenotype. Also expressed at high levels in the cerebellum.
  • Involvement in disease

    Defects in MEIS1 could be a cause of susceptibility to restless legs syndrome type 7 (RLS7) [MIM:612853]. Restless legs syndrome (RLS) is a neurologic sleep/wake disorder characterized by uncomfortable and unpleasant sensations in the legs that appear at rest, usually at night, inducing an irresistible desire to move the legs. The disorder results in nocturnal insomnia and chronic sleep deprivation.
  • Sequence similarities

    Belongs to the TALE/MEIS homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Homeo box protein Meis1 antibody
    • Homeobox protein Meis1 antibody
    • Leukemogenic homolog protein antibody
    • MEIS 1 antibody
    • Meis homeo box 1 antibody
    • Meis homeobox 1 antibody
    • Meis1 antibody
    • Meis1 mouse homolog antibody
    • Meis1 myeloid ecotropic viral integration site 1 homolog antibody
    • Meis1 myeloid ecotropic viral integration site 1 homolog mouse antibody
    • MEIS1 protein antibody
    • MEIS1_HUMAN antibody
    • MGC43380 antibody
    • Myeloid ecotropic viral integration site 1 homolog antibody
    • WUGSC:H NH0444B04.1 antibody
    see all

Images

  • IHC image of MEIS1 staining in Rat normal brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab19867, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Anti-MEIS1 antibody - ChIP Grade (ab19867) at 1 µg/ml (blocked with 5% milk) + KG-1 (Human myeloid cell line) Nuclear Lysate at 15 µg

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

    Performed under reducing conditions.

    Predicted band size: 43, 50 kDa
    Observed band size: 40,48 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes


    This antibody was raised against an immunogen that is predicted to recognize isoforms 1 and 2 of MEIS1. The predicted molecular weights of isoforms 1 and 2 are 43 kDa and 50 kDa respectively. Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • Anti-MEIS1 antibody - ChIP Grade (ab19867) at 1 µg/ml + HEK293 Wcl Transiently Overexpressing MEIS1 at 5 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 43, 50 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:

  • Sanz-Navarro M  et al. Dental Epithelial Stem Cells Express the Developmental Regulator Meis1. Front Physiol 10:249 (2019). Read more (PubMed: 30914971) »
  • Dupays L  et al. Furin, a transcriptional target of NKX2-5, has an essential role in heart development and function. PLoS One 14:e0212992 (2019). Read more (PubMed: 30840660) »
See all 20 Publications for this product

Customer reviews and Q&As

1-3 of 3 Q&A

Question

Good morning,

I filled out the questions the best I could, I hope you can find some conditions I could improve.

Thank you very much for your time,


Order Details
Antibody code: ab19867

Problem
Choose: Non-specific band Multiple bands (and the correct one is missing, so I guess that also makes it weak signal?)


Lot number
GR76623-1


General Information
Antibody storage conditions (temperature/reconstitution etc)
AB reconstituted in 100% glycerol, stored at -20

Description of the problem (high background, wrong band size, more bands, no band etc.)
Expected band (ca 43kDa according to data sheet) not seen; persistant band at ca 60kDa

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
cell extract from human CRC cell lines (LS180 and HCT116; HCT116 is known to have no MEIS1 RNA, and thus not expected to have MEIS1 protein, LS180 does express MEIS1.)

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
10%SDS with complete inhibitor and PHOSstop (Roche), samples heated 5min at 100 degrees after isolation. Re-heated just before loading (and adding of leamli buffer)

Amount of protein loaded
10ug

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
10% bisacrylamide with 0,1% SDS

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
blot buffer : Bio-rad 161-0771. transfer for 1 hr at 100V; largest proteins on marker (250kDa did transfer, so no reason to expect MEIS1 not to be transferred)

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
MEIS1 Rb polyclonal, diluted in TBS-T (0,05%Tween) +5% BSA/dried milk

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Gt anti Rb ::HRPO 1:10.000 in TBST, 1hr at rt, washed 4x for 15 min in TBST lightly shaking

Detection method (ECL, ECLPlus etc.)
ECL

Positive and negative controls used (please specify)
non-primary antibody control (in the picture columns A1 and B1)

No positive control taken along, but same prot. Lysates were used for other blots, and showed no signs of degradation.

Optimization attempts (problem solving)
How many times have you tried the Western?
3x 2blots

Conditions tested:

BSA vs dried milk (both at 5%) (=first test, detected with ECL and incubated with primary AB for 1 hr at RT)

Showed less signal then the 3rd test, but patern was similar

Nitrocellulose membrane with odyssey labeling as detection method (=second test, primary AB for 1hr at RT)

Only 60kDa band was visible for the 1:2000 and 1:4000, but very faint, and for 1:1000 many bands appeared

Primary AB O/N at RT (=3rd test, of which the picture is included; detection with ECL, membrane = PVDF

See result in included picture, longer AB incubation results in more aspecific bands; and no band at the expected size.


Have you run a "No Primary" control?
Yes

Do you obtain the same results every time?
Yes
e.g. are the background bands always in the same place?
60kDa band always appears

What steps have you altered?
membrane (PVDF vs nitrocellulose, detection method; odyssey (using ALEXA-labeled secondary AB) vs ECL, primary AB incubation for 1hr or O/N

Additional Notes:
enclosed picture is with ECL with 30 seconds exposure; PVDF membrane, o/n incubation with primary AB at RT,

Read More
Answer

Thank you for taking the time to complete our questionnaire and contact us. 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 am sorry I am not able to trace this order number on our system, and there are no orders from the Netherlands in March or April this year. Please confirm if the product code is correct (ab19867 Anti-MEIS1 antibody - ChIP Grade) and please provide the name and address of the institution where the order was placed from. If you ordered from a purchasing agent somewhere else, then they should have the Abcam order reference number.

2. We recommend to store antibodies as directed on the datasheet so that they will remain stable and that we can continue to provide our guarantee. If addition of glycerol is required, this will be mentioned on the datasheet. For future reference, this antibody should be aliquotted and stored as it is, as directed on the datasheet. Extra glycerol is not required in this case. Storing an antibody at lower concentrations can affect its stability.

3. I can recommend to lyse the cells in RIPA lysis buffer, which should provide a more suitable protein preparation. Once the lysate is prepared, you can then add sample buffer including reducing and denaturing agents, and heat.

4. What dilutions of antibody have been tried? To reduce the background, I c an recommend to try reducing the concentration of antibody. Incubate overnight at 4oC, which can provide more specific and efficient staining.

5. I can recommend to add a wash step after the primary antibody incubation which will help to wash away any excess antibody.

6. Could you confirm the current vial of secondary antibody is working well with other primary antibodies? I suggest it would be beneficial to include a no primary control to assess if the secondary antibody is binding non specifically. It may be that the concentration of secondary antibody needs to be reduced in order to help optimize the results.

7. We recommend not to mix blocking agents in an experiment, it can sometimesaffect the results. Try 5% BSA only.

8. Has the quality of the sample and transfer to the membrane been assessed using a loading control?

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 for WB and Mouse, Human and Zebrafish 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,it would be helpful tohave some further information and 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 you could also provide an image 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

Question
Answer

DISCOUNT CODE: ****
Expiration date: DD MM YYYY

I am very pleased to hear you would like to accept our offer and test ab19867 in Axololt. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for Axololt and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://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.

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