Overview

  • Product name
  • Description
    Rabbit polyclonal to SLBP
  • Host species
    Rabbit
  • Specificity
    This antibody reacts with SLBP.
  • Tested applications
    Suitable for: ELISA, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Drosophila melanogaster, Zebrafish
  • Immunogen

    A region within synthetic peptide: INYGKNTIAY DRYIKEVPRH LRQPGIHPKT PNKFKKYSRR SWDQQIKLWK, corresponding to amino acids 142-191 of Human SLBP (Peptide available as ab108380.)

  • Positive control
    • HepG2 cell lysate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituents: 2% Sucrose, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab56027 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
ELISA 1/312500.
IHC-P 1/100.
WB Use a concentration of 0.25 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).Can be blocked with SLBP peptide (ab108380). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function
    RNA-binding protein involved in the histone pre-mRNA processing. Binds the stem-loop structure of replication-dependent histone pre-mRNAs and contributes to efficient 3'-end processing by stabilizing the complex between histone pre-mRNA and U7 small nuclear ribonucleoprotein (snRNP), via the histone downstream element (HDE). Plays an important role in targeting mature histone mRNA from the nucleus to the cytoplasm and to the translation machinery. Stabilizes mature histone mRNA and could be involved in cell-cycle regulation of histone gene expression. Involved in the mechanism by which growing oocytes accumulate histone proteins that support early embryogenesis. Binds to the 5' side of the stem-loop structure of histone pre-mRNAs.
  • Tissue specificity
    Widely expressed.
  • Sequence similarities
    Belongs to the SLBP family.
  • Developmental stage
    Regulated during the cell cycle: protein levels increase 10 to 20 fold in the late G1 and decrease at the S/G2 border.
  • Domain
    Amino acids 31-34, 96-99 and 241-244 are necessary for interaction with the Importin alpha/Importin beta receptor. The first 18 amino acids, amino acids 69-76 and 179-182 are necessary for interaction with TNPO3. Amino acids 31-34, 96-99 and 241-244 are necessary for nuclear localization.
  • Post-translational
    modifications
    Phosphorylated on Thr-61 and Thr-62 in the S-phase. Phosphorylation of Thr-62 by CDK1 primes phosphorylation of Thr-61 by CK2. Phosphorylation of Thr-62 is required for its degradation by the proteasome at the end of the S phase. Its degradation is not required for histone mRNA degradation at the end of the S phase. All the phosphorylated forms detected are present in the cytoplasm. Both unphosphorylated and phosphorylated forms bind the stem-loop structure of histone mRNAs.
  • Cellular localization
    Cytoplasm. Nucleus. Polyribosome-associated. Localizes predominantly in the nucleus at the G1/G2 phases and the beginning of S phase. Through the S phase, partially redistributes to the cytoplasm. Binding to histone mRNA is necessary for cytoplasmic localization. Shuttles between the nucleus and the cytoplasm. Imported in the nucleus by the Importin alpha/Importin beta receptor.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • Hairpin binding protein histone antibody
    • HBP antibody
    • Histone binding protein antibody
    • Histone RNA hairpin binding protein antibody
    • Histone RNA hairpin-binding protein antibody
    • Histone stem-loop-binding protein antibody
    • SLBP antibody
    • SLBP_HUMAN antibody
    • Stem loop (histone) binding protein antibody
    • Stem loop binding protein antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bronchial epithelial tissue labelling SLBP with ab56027 at 1/100. A Cy3-conjugated donkey anti-rabbit IgG (1/200) was used as teh secondary antibody. Positive staining shown in the cytoplasm. Magnification: 20X. Exposure time: 0.5 - 2.0 seconds. Left - DAPI. Middle - SLBP. Right - Merge.
  • Anti-SLBP antibody (ab56027) at 0.25 µg/ml + HepG2 cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 30 kDa
    Observed band size: 30 kDa

References

ab56027 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

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 like to offer some suggestions to help optimize the results from ab56027 Anti-SLBP antibody. I would also appreciate if you can confirm some further details:

1. As positive control, I would suggest to use HeLa cells which are treated with Colchicine. This will ensure that you have enough SLBPin you control as theS phase will be prevented. Regarding the negative control, how was the knock down accomplished?

2. As you want to detect protein and not DNA, please calculate the loading so, that you will have a total protein load of 20 to 30 ug per lane.

3. Another method to erase side bands and to amplify proteins of a certain compartment is the use of a nuclear fraction protocol. This allows you as well to measure thequantity of the protein in the nucleus and in the cytoplasm. I have attached the protocol for your convenience.


4. To increase the specificity, please use an antibodydilution of 1/4000.


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for your message.

Would you also please complete the Western blot form sent to you previously? In case you did not get it, I will attach the Word the document again.

Could you confirm what the lysyis buffer contained?





High background and non-specific signal could be due to the secondary antibody. If you have not run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not, I would advise you to check it.



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

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Answer

Thank you for your response.

I have received the Western blot images, thank you very much. However, the completed Questionnaire have not come through yet. Would you be so kind to provide some further information about the Western blot procedure (see attached Word document)? I am particularly interested in the followings:

- sample type (cell, tissue, species),

- sample preparation (lysis buffer used, total lysate or cellular fraction),

- specification of the secondary antibody,

- no primary control,

- positive control

I look forward to hearing from you soon and resolving this issue as soon as possible.

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Question
Answer

Thank you for your response.

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.

I look forward to seeing your Protocol details.

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Answer

Thank you very for your response.

I am happy to take a look at the protocol if you wish to share some details with me. Even if the Abpromise may have been expired.

Let me know if I can help any further.

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Answer

I am sorry to hear that you have been experiencing problems using ab56027 in the application that you wish.

These two antibodies (ab56027 and ab56329) are from different sources and as their datasheets indicate there are differences i.e. clonality, host species, immunogens, tested applications and concentration. Coudl you please take a look at the product documents for further information.

https://www.abcam.com/ab56027: Anti-SLBP antibody

https://www.abcam.com/ab56329: Anti-SLBP antibody

Regarding the complaint of ab56027 - 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 complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

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.

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Question

our customer has performed further experimentsasyou suggested.Please see attachment and our customer's comments.


I send you pictures of westerns made ​​as your have suggested on HeLa cells treated with colchicine; as a further test of nonspecificity I used 3 different secondary antibodies at different concentrations, however, still I do not see any band (or we detect unspecific bands). SLBP KD line was obtained by infection with a short hairpin construct and the KD of 80% was confirmed by TaqMan. I have not tried on nuclear fractions because of SLBP remains bound to histone messenger in the cytoplasm, then cytoplasmic fraction contains much of it.



Wainting for your reply.

Kind regards



Frabcesca


____________________________________________
Dott.ssa Francesca Beretta PhD
Application Specialist
Prodotti Gianni S.p.A. Research Unit
Via Quintiliano 30 20138 Milan Italy
Phone: +39 025097256Fax: +39 025097276
mailto:fberetta@prodottigianni.com http://www.ricerca.it/

Prodotti Gianni is an ISO 9001:2008 Company

PRODOTTIGIANNI è l’unico distributoreautorizzato abcam® per l’Italia.


Da: technical@abcam.com [mailto:technical@abcam.com]
Inviato: giovedì 17 maggio 2012 11:02
A: Beretta Francesca
Oggetto: Reply from Abcam to your enquiry regarding ab56027 [CCE3773954]


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









Dear Dr Beretta



Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

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 like to offer some suggestions to help optimize the results from ab56027 Anti-SLBP antibody. I would also appreciate if you can confirm some further details:

1. As positive control, I would suggest to use HeLa cells which are treated with Colchicine. This will ensure that you have enough SLBPin you control as theS phase will be prevented. Regarding the negative control, how was the knock down accomplished?

2. As you want to detect protein and not DNA, please calculate the loading so, that you will have a total protein load of 20 to 30 ug per lane.

3. Another method to erase side bands and to amplify proteins of a certain compartment is the use of a nuclear fraction protocol. This allows you as well to measure thequantity of the protein in the nucleus and in the cytoplasm. I have attached the protocol for your convenience.


4. To increase the specificity, please use an antibodydilution of 1/4000.


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.



Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3773954


Best regards,
Cathrin

Cathrin Zeeck
Scientific Support Specialist
Abcam plc
www.abcam.com

* Sie können mich auch gerne auf Deutsch kontaktieren


Your original inquiry to Abcam:






Inquiry: Our PO: 1249 (of 27/04/2012) 1)Antibody storage conditions: -20°C, 5 μl aliquotes 2)Description of the problem: more bands/aspecific signal 3)Sample: Cell lysate from HeLa cells 4)Sample preparation: Lysate prepared in reducing Laemly buffer and boyled 5)Amount of protein loaded:1.6ug up to 100ng of DNA 6)Electrophoresis/Gel conditions: Sds page 15% 7)Transfer and blocking conditions:Transfer: 10 min trans blot turbo (biorad) standard protocol rt. Blocking: 1 hour in tbs-t 5% milk rt 8)Primary Antibody:Abcam, rabbit 1:2000/ 1:1000 over night in tbs-t 5%milk +4°, 3 washes in tbs-t 9)Secondary Antibody: GE healthcare, goat anti rabbit Cy5 in tbs-t 1:3000 1 hour rt, 2 washes in tbs-t, 1 wash in pbs Lifetechnologies, goat anti rabbit IgG perox in tbs-t 5%milk 1:3000 1 hour rt, 3 washes in tbs-t 10)Detection method:Typhoon analysis, ECL 11)Positive and negative controls used: Wt hela cells and hela KD for SLBP (20% of mRNA expression) 12)How many times have you tried the Western?: 3 13)Have you run a "No Primary" control?: No but the secondary is proved highly specific using other primary ab 14)Do you obtain the same results every time?:YES 15)What steps have you altered?: Increased the concentration of the ab to check if the signal was too low and lost in the background








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Answer

Thank you for your message and for providing 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 a free of charge replacement or credit note in compensation.

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

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