Overview

  • Product name
  • Description
    Rabbit polyclonal to HEY1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Guinea pig, Cat, Dog
  • Immunogen

    Synthetic peptide within Human HEY1 aa 197-246 (C terminal). The exact sequence is proprietary.
    Sequence:

    PQNGHGNAGTTASPTEPHHQGRLGSAHPEAPALRAPPSGSFGPVLPVVTS


    Database link: Q9Y5J3
    (Peptide available as ab111660)

  • Positive control
    • Jurkat cells, Fetal lung lysate, Human lung or Human kidney tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab22614 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-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 34 kDa.Can be blocked with Human HEY1 peptide (ab111660). 5% skim milk / PBS buffer, and HRP conjugated anti-Rabbit IgG should be diluted in 1/50,000 - 100,000 as second antibody. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use at an assay dependent concentration.

Target

  • Function
    Downstream effector of Notch signaling which may be required for cardiovascular development. Transcriptional repressor which binds preferentially to the canonical E box sequence 5'-CACGTG-3'. Represses transcription by the cardiac transcriptional activators GATA4 and GATA6.
  • Tissue specificity
    Expressed in the somitic mesoderm, the central nervous system, the kidney, the heart, nasal epithelium, and limbs.
  • Sequence similarities
    Belongs to the HEY family.
    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 Orange domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Basic helix loop helix protein OAF1 antibody
    • bHLHb31 antibody
    • Cardiovascular helix loop-helix factor 2 antibody
    • Cardiovascular helix-loop-helix factor 2 antibody
    • CHF-2 antibody
    • CHF2 antibody
    • Class B basic helix-loop-helix protein 31 antibody
    • Hairy and enhancer of split related protein 1 antibody
    • Hairy and enhancer of split-related protein 1 antibody
    • Hairy enhancer-of-split related with YRPW motif protein 1 antibody
    • Hairy related transcription factor 1 antibody
    • Hairy-related transcription factor 1 antibody
    • Hairy/enhancer of split related with YRPW motif 1 antibody
    • Hairy/enhancer-of-split related with YRPW motif protein 1 antibody
    • HERP2 antibody
    • HES related repressor protein 1 antibody
    • HES-related repressor protein 1 antibody
    • HES-related repressor protein 2 antibody
    • HESR-1 antibody
    • HESR1 antibody
    • HEY 1 antibody
    • hey1 antibody
    • HEY1_HUMAN antibody
    • hHRT1 antibody
    • HRT-1 antibody
    • HRT1 antibody
    • OAF1 antibody
    see all

Images

  • Predicted band size: 34 kDa

  • Lane 1 : Anti-HEY1 antibody (ab22614) at 5 µg/ml
    Lane 2 : Anti-HEY1 antibody (ab22614) at 2.5 µg/ml
    Lane 3 : Anti-HEY1 antibody (ab22614) at 1.25 µg/ml

    All lanes : Cell lysate prepared from human fetal lung cells

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 34 kDa

  • Paraffin embedded human lung tissue labelled with HEY1 antibody (ab22614) at 4-8µg/ml.  Magnification 400X.

    Alveolar cells are positively labelled (indicated with arrows).

  • ab14404 staining HEY1 in the COS1 fibroblast cell line from Monkey kidney by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% in PBS and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 16 hour at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal(1/500) was used as the secondary antibody.

    See Abreview

  • Paraffin embeded human kidney tissue labelled with HEY1 antibody (ab22614) at 4-8µg/ml. Magnification 400X.

    Epithelial cells of renal tubules are positively labelled (indicated with arrows).

References

This product has been referenced in:
  • Yin X  et al. Hey1 functions as a positive regulator of odontogenic differentiation in odontoblast-lineage cells. Int J Mol Med 41:331-339 (2018). Read more (PubMed: 29138798) »
  • Cook N  et al. A phase I trial of the ?-secretase inhibitor MK-0752 in combination with gemcitabine in patients with pancreatic ductal adenocarcinoma. Br J Cancer 118:793-801 (2018). IHC-P ; Human . Read more (PubMed: 29438372) »
See all 34 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Skin fibroblast)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
Skin fibroblast
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 28 2017

Abreviews
Application
ChIP
Sample
Mouse Cell lysate - nuclear (SN4741 cells)
Negative control
igG
Specification
SN4741 cells
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde

Abcam user community

Verified customer

Submitted Feb 09 2016

Application
Western blot
Sample
Human Purified protein (brain cancer)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
brain cancer
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 26°C

Abcam user community

Verified customer

Submitted Apr 24 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Placenta)
Specification
Placenta
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Mukesh Jaiswal

Verified customer

Submitted May 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH3T3)
Specification
NIH3T3
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Mar 22 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (COS1 cell line)
Specification
COS1 cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 11 2013

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 (postnatal day 1 testes)
Specification
postnatal day 1 testes
Fixative
4% PFA
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Qing Wen

Verified customer

Submitted Jan 02 2013

Answer

Thank you for contacting us.

I have been in touch with the lab regarding your inquiry. The sample type used in the western blot and IHC images are of normal human lung.The sample type used in the other IHC image is of normal human kidney.

While we can recommend these tissues as control, the gold standard is a knock-out vs. wt in a tissue known to express the protein, or, a knock-in vs. WT in a tissue known not to express the protein. Other IHC controls could be transfected vs. untransfected tissue culture cells.

This product is covered by our Abpromise guarantee. We are happy provide scientific support at any time. If you are using the product in species and applications listed on the datasheet and contact us within six months of purchase, we are also happy to replace or refund the product should we not be able to help you to resolve the issue. More information on our Abpromise may be found at the following link:

https://www.abcam.com/index.html?pageconfig=abpromise

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More

Question
Answer

Thank you for calling Abcam earlier today.

I am sorry that you are experiencing issues with ab22614 in IHC-P. As we discussed on the telephone, if you could either send me the Abcam order number or the PO# that was used to purchase the antibody, I will be able to send a replacement for you to try.

I look forward to your reply.

Read More

Question

Dear technical support:

This customer raised an inquiry about ab22614 (Anti-HEY1 antibody), and this customer used this antibody conduct the western blotassay with human sample, and the data shows no band, because this antibody is already out of our warranty period, therefore she only wants to ask for your kindly help to offer any suggestion to her. I attach the WB questionnaire in this letter, could you please help this customer to solve this problem.

Thanks for your kindly assistance.

Best regards

1. Order details:

Batch number: GR20993-4

Po: 968560

Abcam product code: ab22614

Antibody storage conditions (temperature/reconstitution etc) : -20 degrees

2. Please describe the problem (high background, wrong band size, more bands, no band etc).

No band

3. On what material are you testing the antibody in WB?

· Species: human

· What’s cell line or tissue: cell line

Huh7 / Hep3B / HepG2 / Ha22T / SKHEP1 /HCC26 / Mahlavue

· Cell extract or Nuclear extract: cell extract

· Purified protein or Recombinant protein: purified protein

3. The lysate


How much protein was loaded:

What lysis buffer was used: RIPA lysis buffer

What protease inhibitors were used: PMSF

What loading buffer was used: 5x SDS loading dye

Phosphatase inhibitors : no

Did you heat the samples: temperature and time: 98 degrees / 10min

4. Electrophoresis/Gel conditions/ Transfer conditions

Reducing or non reducing gel: reducing gel

Reducing agent: TEMED

Gel percentage : 10 % PAGE

Transfer conditions: (Type of membrane, Protein transfer verified):

Wet transfer / 110v 90min limit:0.33amp / PVDF transfer membrane

5. Blocking conditions

Buffer: 1X TBST

Blocking agent: milk, BSA, serum, what percentage: 5% milk

Incubation time: 60 min`

Incubation temperature: room temperature

6. Primary Antibody

Species: rabbit

Reacts against:anti-HEY1

· At what dilution(s) have you tested this antibody: 100x/200x/500x

· What dilution buffer was used: TBST

· Incubation time: over night

· Incubation temperature: 4 degrees

· What washing steps were done: TBST wash 10 min / 3 times

7. Secondary Antibody

Species: goat

Reacts against: anti-rabbit IgG HRP

At what dilution(s) have you tested this antibody: 10000x

Incubation time: RT

Wash steps: TBST wash 10min / 3 times

Fluorochrome or enzyme conjugate: HRP conjugated

Do you know whether the problems you are experiencing come from the secondary? I am sure that secondary Ab does work.

8. Detection method
ECl, ECl+, other detection method: ECL

9. Did you apply positive and negative controls along with the samples? Please specify.

Positive control : alpha-tubulin

10. Optimization attempts

· How many times have you tried the Western? 3 times

· Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):

I am sure that secondary Ab works.

· Do you obtain the same results every time e.g. are background bands always in the same place? Repeated the same results ( No band)

· What steps have you altered? Decrease Ab dilution -> No band

Read More
Answer

Thank you for taking time to complete our questionnaire and informing us of the problems that your customer has been having with this antibody.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would also appreciate it if youcould confirm some further details:

1. You said that both whole cell extract and recombinant protein was used to perform the blot? How was the recombinant protein produced? Is it possible to share an image of the blot obtained, along with that produced with the alpha-tubulin control?

2. How much protein was loaded in each well?

3. Has the transfer been verified by staining of the membrane?

4. Was thealpha-tubulinprobed on the same membrane?If so was a stripping procedure employed? Which protein target was probed first?

5. How long was the membraneexposed for?

Once I have this additional information I will be able to give the customer a few suggestions which may help to improve the results observed so far.

I thank you for your cooperation and look forward to your reply.

Read More

1-10 of 12 Abreviews or Q&A

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

Sign up