Overview

  • Product name

  • Description

    Rabbit polyclonal to Twist
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Chicken, Cow, Cat, Dog, Pig, Chimpanzee, Zebrafish
  • Immunogen

    Synthetic peptide within Human Twist aa 141-190 (C terminal). The exact sequence is proprietary.
    Sequence:

    DKLSKIQTLKLAARYIDFLYQVLQSDELDSKMASCSYVAHERLSYAFSVW


    Database link: Q15672
    (Peptide available as ab111667)

  • Positive control

    • Jurkat whole cell lysate (ab7899).

Properties

Applications

Our Abpromise guarantee covers the use of ab49254 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 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 2.5 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 21 kDa).

Abcam recommends blocking with BSA.

Target

  • Function

    Acts as a transcriptional regulator. Inhibits myogenesis by sequestrating E proteins, inhibiting trans-activation by MEF2, and inhibiting DNA-binding by MYOD1 through physical interaction. This interaction probably involves the basic domains of both proteins. Also represses expression of proinflammatory cytokines such as TNFA and IL1B. Regulates cranial suture patterning and fusion. Activates transcription as a heterodimer with E proteins. Regulates gene expression differentially, depending on dimer composition. Homodimers induce expression of FGFR2 and POSTN while heterodimers repress FGFR2 and POSTN expression and induce THBS1 expression. Heterodimerization is also required for osteoblast differentiation.
  • Tissue specificity

    Subset of mesodermal cells.
  • Involvement in disease

    Defects in TWIST1 are a cause of Saethre-Chotzen syndrome (SCS) [MIM:101400]; also known as acrocephalosyndactyly type 3 (ACS3). SCS is a craniosynostosis syndrome characterized by coronal synostosis, brachycephaly, low frontal hairline, facial asymmetry, hypertelorism, broad halluces, and clinodactyly.
    Defects in TWIST1 are the cause of Robinow-Sorauf syndrome (RSS) [MIM:180750]; also known as craniosynostosis-bifid hallux syndrome. RSS is an autosomal dominant defect characterized by minor skull and limb anomalies which is very similar to Saethre-Chotzen syndrome.
    Defects in TWIST1 are the cause of craniosynostosis type 1 (CRS1) [MIM:123100]. Craniosynostosis consists of premature fusion of one or more cranial sutures, resulting in an abnormal head shape.
  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • ACS3 antibody
    • B-HLH DNA binding protein antibody
    • bHLHa38 antibody
    • BPES2 antibody
    • BPES3 antibody
    • Class A basic helix-loop-helix protein 38 antibody
    • CRS antibody
    • CRS1 antibody
    • CSO antibody
    • H-twist antibody
    • OTTHUMP00000116043 antibody
    • SCS antibody
    • Twist basic helix loop helix transcription factor 1 antibody
    • Twist family bHLH transcription factor 1 antibody
    • Twist homolog 1 (Drosophila) antibody
    • Twist homolog 1 antibody
    • TWIST homolog of drosophila antibody
    • Twist related protein 1 antibody
    • Twist-related protein 1 antibody
    • TWIST1 antibody
    • TWST1_HUMAN antibody
    see all

Images

  • Anti-Twist antibody (ab49254) at 2.5 µg/ml + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50,000 - 1/100,000

    Predicted band size: 21 kDa
    Observed band size: 22 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemistry of Uterus tissue at an antibody concentration of 4-8µg/ml using anti-TWIST1 antibody (ab49254)

  • ab49254 staining Twist in mouse lung adencarcinoma induced by urethane tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then blocked with 4% serum for 2 hours at 25°C followed by incubation with the primary antibody, at a 1/100 dilution, for 16 hours at 4°C. A biotinylated goat anti-rabbit IgG polyclonal was used as secondary antibody at a 1/200 dilution.

    See Abreview

  • ab49254 staining Twist in human testis tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer, pH 6.0 prior to blocking with 10% serum for 1 hour at 20°C. The primary antibody was diluted 1/50 and incubated with the sample for 12 hour at 4°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody, diluted 1/200.

    See Abreview

  • All lanes : Anti-Twist antibody (ab49254)

    Lane 1 : Jurkat cell lysate
    Lane 2 : Jurkat cell lysate with blocking peptide at 2 µg/ml

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 21 kDa

References

This product has been referenced in:

  • Shi J  et al. NUP58 facilitates metastasis and epithelial-mesenchymal transition of lung adenocarcinoma via the GSK-3ß/Snail signaling pathway. Am J Transl Res 11:393-405 (2019). Read more (PubMed: 30787996) »
  • Pang K  et al. The ERH gene regulates migration and invasion in 5637 and T24 bladder cancer cells. BMC Cancer 19:225 (2019). Read more (PubMed: 30866868) »
See all 37 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Answer

Thank you for contacting us.

I can confirm that the recommended concentration for this product in Western Blot is 2.5ug/ml. I have also reviewed the publication listed on the datasheet and found that they had used a 2.0ug/ml concentration of this product in Western Blot. While these concentrations are good starting points for use, we do recommend that you do run a dilution series with any new antibody to titer the optimal dilution for your protocol.

You may also be able reduce the amount of solution used in your antibody incubation steps by incubating in a sealed bag, we recommend incubating 1ml of primary antibody solution per 15 cm2 blot.Hybridization tubes or 50ml Falcon tubes can also be used (˜2.5ml primary antibody/blot) may also be used.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (ES cells)
Gel Running Conditions
Reduced Denaturing (Bis-Tris from Invitrogen 4-12%)
Loading amount
18 µg
Specification
ES cells
Blocking step
BSA as blocking agent for 5 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Maud de Dieuleveult

Verified customer

Submitted Jul 21 2017

Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3)
Gel Running Conditions
Reduced Denaturing (12% Bis tris)
Loading amount
50 µg
Specification
3T3
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 30 2015

Application
IHC - Wholemount
Sample
Zebrafish Embryo (wholemount)
Specification
wholemount

Abcam user community

Verified customer

Submitted Dec 04 2013

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (gel 11%)
Sample
Human Cell lysate - whole cell (ovarian cancer cell line SKOV3)
Specification
ovarian cancer cell line SKOV3
Blocking step
blocking One (Nacalai Tesque) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C

Dr. Senn Wakahashi

Verified customer

Submitted Jul 08 2013

Answer

Thank you for your updates.

I am very sorry to hear that the customer is not satisfied with the staining.

I can certainly offer either a new vial as a free of charge replacement or a credit note which can be used for future purchase or for a full reimbursement.

Could you please discuss it with your customer and let me know how you wish to proceed.

I look forward to hearing from you soon.

Read More

Question

Dear Sir,

About the sample type, we have used formalin fixed paraffin embedded sections of human normal testis as in the data sheet and another case of colon carcinoma.

the method used was streptavidin-biotin amplified system. The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval

in citric buffer, pH 6.0 for 25min prior to blocking with hydrogen peroxide.The primary

antibody was diluted 1/50 and incubated with the sample for 16 hour .An HRP-

conjugated goat anti-rabbit polyclonal was used as the secondary antibody, diluted

1/200.

-Washing was done using phosphate buffer saline (PBS)

-The detection Kit is Ultra Vision LP detection system anti-poly valent HRP/DAB (catalog #TP-015-HD) (Lab vision, USA). In this system, two reagents were

utilized:

The biotinlated secondary anti-immunoglobulin which is a purified goat polyvalent anti-mouse IgG capable of binding to both the primary antibody. ü

The streptavidin-biotin enzyme complex. ü

Finally, the reaction was visualized by substrate/chromogen Diaminobenzidine, DAB) reagent.

- the controls used is human normal testis tissue as in the datasheet.














Sincerely yours,

Ayman Sadek
Kemet Medical
M: +20100 5094 222
F: +20226903126
O: +20226903127 / +20226903128
E: mailto:asadek@kemet.com.eg
A: 14a El Emam Aly St.
Heliopolis Cairo Egypt 11341
PO Box 158 Heliopolis














From: technical@abcam.com [mailto:technical@abcam.com]
Sent: Tuesday, February 26, 2013 1:27 PM
To: Ayman S. Abdel Rehim
Subject: Reply from Abcam to your enquiry regarding ab49254 [CCE4702003]



















https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header









Dear Ayman




I am sorry to hear that your customer has been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Could you please provide some further details of the protocol used and further information see the following points below. It would be much appreciated if you could attach an image to the response.

Immunohistochemistry/Immunocytochemistry


Sample type, species, tissue

Sample fixation, antigen retrieval, permeabilization

Blocking conditions

Primary antibody conditions (diluent/dilution, incubation time/temperature)

Secondary antibody conditions (diluent/dilution, incubation time/temperature)

Wash steps

Detection method, amplification steps (if any)

Controls used (sample, alternative primary ab, isotype control)

Lot number/order number/date of purchase


Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Best regards,
Tanya

Tanya Bagrij, PhD
Scientific Support Supervisor
Abcam plc
www.abcam.com




Your original inquiry to Abcam:














Dear Sir,
We recently bought your antibody Ab49254 – an anti-twist antibody, rabbit polyclonal antibody. We have tested the antibody on histological sections of the positive control in datasheet, normal human testis and on colon carcinoma two times. We have used paraffin embedded tissue that have pretreated paraffin-embedded in citrate buffer pH 6.0 and heated in a microwave oven for25min for antigen retrieval. we have proceeded in the streptavidin biotin technique as beeing used for IHC using DAB as counter stain and myers heamatoxylin. In all cases the antibody result in a cytoplasmic staining with no indication of nuclear staining as expected and been told in the data sheet. please help me solving this problem as i will only be able to deel with the nuclear results in my reasearch.
waiting for reply.






























































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Read More
Answer

Thank you for your response.

After reading through the detailed protocol you kindly forwarded to Abcam, I would make some comments:

1) Image: Could you please save the image as a jpeg file and send it back to us.

2) Antigen retriaval: heat-mediated antigen retrieval in citric buffer, pH 6.0 for 25min prior

25 mins seems to be very long. Did the customer use microwave, waterbath or vegetable steamer for antigen retrieval? Please check this?

Three minutes is only suggested as a starting point antigen retrieval time. Less than 3 minutes may leave the antigens under-retrieved, leading to weak staining. More than 3 minutes may leave them overretrieved, leading to non-specific background staining and also increasing the chances of sections dissociating from the slides. A control experiment is recommended beforehand, where slides of the same tissue section are retrieved for 1, 2, 3, 4 and 5 minutes before being immunohistochemically stained to evaluate optimum antigen retrieval time for the particular antibody being used.

3) Incubation with the primary antibody:

Since signal enhancing system was used, 16 hrs incubation with the primary antibody seems to be very long and it may contribute to non-specific signal. I would recommend using this antibody at a dilution of 1/100 for 1 or 2 hrs at room temperature (25 oC).

4) Detection system:

Does the detection system work fine? Have you used it successfully with another primary antibody?

5) Blocking:

There is no information about any blocking step applied in the e-mail. Please make sure that non-specific binding sites are blocked either with 5% BSA or 10% serum (one hour at room temperature).

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (NMuMG)
Loading amount
75 µg
Specification
NMuMG
Gel Running Conditions
Reduced Denaturing (12)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 19 2012

Answer

Thank you for kindly confirming these details as this enables us to closely monitor the quality of our products

I would be pleased to arrange a replacement product free of charge. I have added a vial of ab50887 to your next order.


I hope this information is helpful and satisfactory for the customer. Please do not hesitate to contact us if you need anything further.

Read More

Answer

Thank you for taking time to provide that information and for contacting us with this problem. I am sorry to hear this antibody hasnotbeen providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been more successful.I apologize for the inconvenience this has caused you and I would bepleased to offer you a free of charge replacement with ab50581 (there is no need to pay the difference).

In order to arrange this, could you please provide me with the order number of ab49254 (or the order date and delivery address).

Thank you for your cooperation. I look forward to receiving your reply.

Read More

1-10 of 19 Abreviews or Q&A

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