Overview

  • Product name
    Anti-BLBP antibody - Neuronal Marker
    See all BLBP primary antibodies
  • Description
    Rabbit polyclonal to BLBP - Neuronal Marker
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, IHC-P, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Zebrafish, Quail
    Predicted to work with: Human, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse BLBP.

    Read Abcam's proprietary immunogen policy (Peptide available as ab32422.)

  • Positive control
    • Rat spinal cord from embryonic day 16; young zebrafish spinal cord This antibody gave a positive signal in the following mouse lysate: Brain (Mouse) Tissue Lysate - normal tissue, 0 days old

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32423 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-FoFr 1/2000.
IHC-P 1/200 - 1/1000.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).Can be blocked with Mouse BLBP peptide (ab32422).

Abcam recommends using milk as the blocking agent.

IHC-Fr Use at an assay dependent concentration. PubMed: 23675446

Target

  • Function
    B-FABP could be involved in the transport of a so far unknown hydrophobic ligand with potential morphogenic activity during CNS development. It is required for the establishment of the radial glial fiber system in developing brain, a system that is necessary for the migration of immature neurons to establish cortical layers.
  • Tissue specificity
    Expressed in brain and other neural tissues.
  • Sequence similarities
    Belongs to the calycin superfamily. Fatty-acid binding protein (FABP) family.
  • Domain
    Forms a beta-barrel structure that accommodates the hydrophobic ligand in its interior.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • B FABP antibody
    • B-FABP antibody
    • BFABP antibody
    • BLBP antibody
    • Brain lipid binding protein antibody
    • Brain lipid-binding protein antibody
    • Brain-type fatty acid-binding protein antibody
    • DKFZp547J2313 antibody
    • FABP 7 antibody
    • FABP7 antibody
    • FABP7_HUMAN antibody
    • FABPB antibody
    • Fatty Acid Binding Protein 7 antibody
    • Fatty acid binding protein 7 brain antibody
    • Fatty acid binding protein brain antibody
    • Fatty acid-binding protein 7 antibody
    • Fatty acid-binding protein, brain antibody
    • Mammary derived growth inhibitor related antibody
    • Mammary-derived growth inhibitor related antibody
    • MRG antibody
    • OTTHUMP00000017119 antibody
    see all

Images

  • BLBP detected on rat embryo day 16 spinal cord T/S on 5 micron thick formalin-fixed paraffin wax embedded sections using Rabbit polyclonal to BLBP (ab32423; incubated at 1/200 for 16h at RT). 1% BSA (10 mins at RT) used as blocking agent. Secondary Antibody was biotin labeled anti-rabbit IgG (1/300). Immunoreactivity was visualised by standard streptAvidin-biotin-complex using peroxidase as a reporter and DAB as chromogen following citric acid (pH 6) heat mediated antigen retrieval. Expected specific staining observed with ab32423 with low signal to noise ratio.
  • IHC-Fr image of Pax2 staining on zebrafish retina sections using ab32423 (1:100). The sections were paraformaldehyde and permeabilized using Triton-X. Antigen retrieval was performed using Tris-HCl and blocking was perfomed using 5% BSA for 1 hour at 23°C. ab32423 was diluted 1:100 and incubated with the sections for 16 hours at 4°C. The secondary antibody was goat polyclonal to rabbit IgG conjugated to Alexa Fluor 488 (1:1000).

    See Abreview

  • Anti-BLBP antibody - Neuronal Marker (ab32423) at 1/250 dilution + Mouse brain tissue lysate - total protein (0 days) (ab7188) at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

  • ICC/IF image of ab32423 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab32423, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • BLBP detected on mouse embryo day 16 spinal cord T/S on 5 micron thick formalin-fixed paraffin wax embedded sections using Rabbit polyclonal to BLBP (ab32423; incubated at 1/300 for 16h at RT). 1% BSA (10 mins at RT) used as blocking agent. Secondary Antibody was biotin labeled anti-rabbit IgG (1/300). Immunoreactivity was visualised by standard streptAvidin-biotin-complex using peroxidase as a reporter and DAB as chromogen following citric acid (pH 6) heat mediated antigen retrieval. Expected specific staining observed with ab32423.
  • Immunohistochemistical staining (Formaldehyde/PFA-fixed paraffin-embedded sections) for BLBP antibody - Neuronal Marker (ab32423)on Zebrafish Tissue sections (young adult whole body T/S). Antigen retrieval step: Heat mediated Blocking step: 1% BSA for 10 mins at RT. Primary Antibody ab32423 incubated at 1/500 for 2 hours at RT. Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).
  • Immunohistochemistical detection (on Formaldehyde /PFA-fixed paraffin-embedded sections) of BLBP protein using BLBP antibody - Neuronal Marker (ab32423) on Quail Tissue sections (developing spinal cord). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody ab32423 incubated at 1/500 for 2 hours at RT. Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).

References

This product has been referenced in:
  • Sagner A  et al. Olig2 and Hes regulatory dynamics during motor neuron differentiation revealed by single cell transcriptomics. PLoS Biol 16:e2003127 (2018). Read more (PubMed: 29389974) »
  • Moreno N & González A Pattern of Neurogenesis and Identification of Neuronal Progenitor Subtypes during Pallial Development in Xenopus laevis. Front Neuroanat 11:24 (2017). Read more (PubMed: 28396626) »
See all 47 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (Retina, Muller glia cells)
Specification
Retina, Muller glia cells
Permeabilization
Yes - triton X
Fixative
Paraformaldehyde

Dr. Ryan Macdonald

Verified customer

Submitted Aug 22 2014

Answer

Muchas gracias por enviarnos el protocolo relleno. Siento mucho que este anticuerpo os este dando problemas en IHC.

Para poder ayudaros a resolver el problema sufrido con el anticuerpo quisiera que me aclararais algunos detalles:

¿No se ha obtenido ninguna tinción en el control positivo? ¿Habéis comprobado la expresión de la proteína en el tejido?
¿Se ha optimizado el proceso de recuperación antigénica probando distintas temperaturas, buffers y tiempos?
¿Podríais por favor facilitarme el numero de lote / pedido de los viales que os funcionaron bien en el pasado?
Os agradecería que por favor me mandarais una imagen de ambos lotes, si está disponible, para poder compararlos.



Me imagino que el resto de productos implicados en el proceso se han verificado (secundario, sustrato cromogénico, etc..).

Como sabéis, todos nuestros productos están cubiertos por la garantía Abpromise, que garantiza que si se han usado siguiendo las especificaciones de la datasheet (especies y aplicaciones indicadas), y el protocolo de utilización de los mismos es adecuado, os mandaremos un reemplazo del vial totalmente gratuito, un reembolso o un vale por el precio del producto, siempre que la compra se haya realizado en los últimos 6 meses.

Así pues, quedo a la espera de que me confirmes estos detalles para poder proceder, y solucionar este incidente.

Muchas gracias por tu colaboración y que pases un buen fin de semana.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary neural culture)
Specification
Primary neural culture
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 21 2012

Answer

Thank you for contacting us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to investigate this particular case further for you, and also obtain more information for our quality records. In order to proceed with this, there are a few things I would like to clarify to better understand and solve the problems experienced with this antibody.

Could you please describe the problem experienced with the antibody? Didn’t you observe any signal at all, unspecific signal, high background? I would appreciate if you could please provide an image of the tissue sections in order to better understand the problem described.

Also, the order number and / or lot number is also important to dismiss shipping failure and monitor our product’s quality. Could you please provide this information?

Regarding the protocol followed, was the antigen retrieved after fixation? If so, which method was used? Was the secondary conjugated to an enzyme or was it a fluorometric method?

Unfortunately, this antibody has only been tested with frozen and paraffin embedded sections, we do not have any evidence to support its performance in agarose embedded sections. In order to assess the antibody’s activity, I would suggest trying a protocol which has been proven to give positive results in our laboratory. If you had frozen or paraffined sections available, I would suggest giving it a try.

Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered.

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

Read More

Answer

Thank you for your note.
First please do try and use the free of charge replacement and let me know how you are getting on with ab32423.
Keep in touch and good luck with the experiments!

Read More

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 1154518.
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 enquiry regarding abab32423 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.
I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that 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.
It seems that the samples were fixed twice i.e. perfusion fixation and post-fixation with (PFA/formalin fixed paraffin embedded sections). Have you tried to apply a gentle antigen retrieval (HIER) to see if the signal is getting stronger?
I could offer you a new vial as a free of charge replacement but we do not have a different batch in stock. So it would be from the same you have just used.
Could you let me know how you wish to proceed with your enquiry. I look forward to hearing from you soon.

Read More

Answer

I am sorry to hear that you have 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 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.
I am particularly interested in the following information:
- sample preparation,
- antigen retrieval,
- positive control used
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.

Read More

Answer

Thank you for contacting us again and for providing some further details.

It may well be that the antibody has been accidentally damaged during shipment or transfer. I would suggest aliquotting this product and storing at -20°C or -80°C.

I could offer a new vial for this troubled customer and would like to ask you to advise him/her to aliquot this antibody and to keep the vials in the freezer.

Please do let me know how you wish to proceed with this enquiry.

Read More

Answer

Thank you for contacting us. Please follow the procedure as given below; - Embed tissue in OCT medium. - Section it using cryostat - Place tissue sections on glass slide and dry the sections overnight at RT. - Fix the sections with suitable fixative acetone or ethanol (-20C) for 10 minutes. This step eliminates the use of antigen retrieval in IHC on frozen sections. In case of formaldehyde fixation there is always a need of epitope retrieval. - Pour off acetone and allow it to evaporate for > 20 min at room temperature. - Rinse the slides phosphate buffered saline (PBS) for 2 changes, 5 min each. - Incubate slides in 0.3% H2O2 to block endogenous peroxidase. - Rinse with PBS - Add blocking buffer - Apply primary and secondary antibody as usual. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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1-10 of 18 Abreviews or Q&A

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