製品の概要

  • 製品名

    Anti-beta III Tubulin antibody
    beta III Tubulin 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to beta III Tubulin
  • 由来種

    Rabbit
  • 特異性

    The immunogen used for this product shares 75% homology with TUB (Tubby protein homolog, Uniprot: P50607). In western blot, we observe a specific band at ~55kDa which is not seen in KO cell lines. An additional band below this band of interest is seen at ~50kDa in both the WT and KO cells which could correspond to the protein TUB. Please note that cross-reactivity with this protein has not been confirmed experimentally. TUB is localized notably in high concentrations in the nucleoli of brain neurons with lower protein levels in the cytoplasm. Please, therefore, be aware that ICC experiments may need to be optimised. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. As an alternative antibody, we would recommend our recombinant rabbit monoclonal antibody ab52623 which has been shown to specific in both WB and ICC using KO cells.
  • アプリケーション

    適用あり: IHC-FoFr, Flow Cyt, IHC - Wholemount, IHC-P, IHC-P, IHC-Fr, ICC/IF, WBmore details
  • 種交差性

    交差種: Mouse, Rat, Human, Pig, Rhesus monkey, Common marmoset, Dogfish, Catshark
  • 免疫原

    Synthetic peptide corresponding to Human beta III Tubulin aa 350 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab18660)

  • ポジティブ・コントロール

    • WB: HAP1 cell lysate; Mouse brain and hippocampus lysates; rat brain lysate; human brain lysate. ICC/IF: SKNSH, Neuro-2A and NGF-differentiated PC12 cells IHC-P: Human cerebellum tissue; mouse brain tissue; rat cerebellum tissue; dogfish/catshark head tissue. Flow cyt: U-87MG and Neuro 2A cells.
  • 特記事項

      

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab18207 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-FoFr Use at an assay dependent concentration. PubMed: 20568963
Flow Cyt Use 0.01µg for 106 cells.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody

IHC - Wholemount Use at an assay dependent concentration. PubMed: 25383879
IHC-P 1/2000.
IHC-P 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.

We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody

WB Use a concentration of 1 µg/ml. Detects a band of approximately 50-55 kDa (predicted molecular weight: 50 kDa).Can be blocked with Human beta III Tubulin peptide (ab18660).

ターゲット情報

  • 機能

    Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha-chain. TUBB3 plays a critical role in proper axon guidance and mantainance.
  • 組織特異性

    Expression is primarily restricted to central and peripheral nervous system.
  • 関連疾患

    Defects in TUBB3 are the cause of congenital fibrosis of extraocular muscles type 3A (CFEOM3A) [MIM:600638]. A congenital ocular motility disorder marked by restrictive ophthalmoplegia affecting extraocular muscles innervated by the oculomotor and/or trochlear nerves. It is clinically characterized by anchoring of the eyes in downward gaze, ptosis, and backward tilt of the head. Congenital fibrosis of extraocular muscles type 3 presents as a non-progressive, autosomal dominant disorder with variable expression. Patients may be bilaterally or unilaterally affected, and their oculo-motility defects range from complete ophthalmoplegia (with the eyes fixed in a hypo- and exotropic position), to mild asymptomatic restrictions of ocular movement. Ptosis, refractive error, amblyopia, and compensatory head positions are associated with the more severe forms of the disorder. In some cases the ocular phenotype is accompanied by additional features including developmental delay, corpus callosum agenesis, basal ganglia dysmorphism, facial weakness, polyneuropathy.
  • 配列類似性

    Belongs to the tubulin family.
  • ドメイン

    The highly acidic C-terminal region may bind cations such as calcium.
  • 翻訳後修飾

    Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
  • 細胞内局在

    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 参照データベース

  • 別名

    • beta 3 tubulin antibody
    • beta 4 antibody
    • beta-4 antibody
    • CDCBM antibody
    • CDCBM1 antibody
    • CFEOM3 antibody
    • CFEOM3A antibody
    • FEOM3 antibody
    • M(beta)3 antibody
    • M(beta)6 antibody
    • MC1R antibody
    • Neuron specific beta III Tubulin antibody
    • Neuron-specific class III beta-tubulin antibody
    • QccE-11995 antibody
    • QccE-15186 antibody
    • TBB3_HUMAN antibody
    • Tubb 3 antibody
    • TUBB3 antibody
    • TUBB4 antibody
    • Tubulin beta 3 antibody
    • Tubulin beta 3 chain antibody
    • Tubulin beta 4 antibody
    • Tubulin beta III antibody
    • Tubulin beta-3 chain antibody
    • Tubulin beta-4 chain antibody
    • Tubulin beta-III antibody
    • tuj 1 antibody
    • tuj1 antibody
    see all

画像

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Beta III Tubulin knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab18207 observed at 55 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab18207 was shown to recognize beta III Tubulin in wild-type HAP1 cells as signal was lost in beta III Tubulin knockout cells. An additional cross-reactive band at 50 kDa was observed in wild-type and knockout cells. Due to the immunogen’s homology with TUB (Tubby protein homolog, Uniprot: P50607), this lower band could correspond to the TUB protein. Please note that cross-reactivity with this protein has not been confirmed experimentally.

    Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. Ab18207 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature

  • ab18207 staining beta III Tubulin in NGF-differentiated PC12 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18207 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
    Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • IHC image of ab18207 staining beta III Tubulin in rat cerebellum formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab18207, 1:2000 dilution, 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. No primary antibody was used in the secondary only control (shown on the inset).
    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.

  • Differential expression of Dnmt1, Dnmt3a, and Dnmt3b during RA induced neuronal differentiation of P19 cells

    Mouse P19 cells either left untreated (top panel) or RA treated for initial 2 days and further cultured for 4 days without RA (6 days, bottom panel) were immunostained with neuron specific β-III tubulin antibody and nuclei were stained using DAPI.

    In order to confirm the neuronal morphology, the cells were stained for neuron specific beta III-tubulin (ab18207). RA induced P19 cells showed immunoreactivity against βIII-tubulin, indicating a neuronal phenotype. In contrast, undifferentiated P19 cells were βIII-tubulin negative.

    (After Figure 1A of Sheikh et al)

  • Immunohistochemical analysis of adult mice ovaries undergone Clarity processing staining tyrosine hydroxlase (TH), Beta III Tubulin (Tuj1) with ab18207, and brain derived neurotrpic factor (BDNF) with ab72439. Positive staing of Tuj1 and BDNF is evident in the theca cells and corpus luteum.

  • Reduction of corneal nerves in CD207-dDTR+DE mice

    After 7 days of DE induction of WT mice and CD207-dDTR mice, immunostaining for beta III tubulin (green) on a corneal flap mount was performed and compared with a non-DE induced control (CONT). High magnification (× 100 upper row and × 200 lower row) images were taken.

    The cornea samples were incubated in diluted primary antibody (ab18207) overnight at 37°C. Corneas were then washed in PBS for one hour, and then washed again. Diluted secondary antibody (FITC anti-rabbit ) was applied to the corneas with overnight shaking at 4°C. After two one-hour washes in PBS, corneas were mounted on glass slides with mounting medium.

  • Overlay histogram showing Neuro 2A cells stained with ab18207 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18207, 0.01μg/1x106) for 30 min at 22ºC. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • All lanes : Anti-beta III Tubulin antibody (ab18207) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Brain (Rat) Tissue Lysate
    Lane 4 : Human brain tissue lysate - total protein (ab29466) with Human beta III Tubulin peptide (ab18660) at 2 µg/ml
    Lane 5 : Brain (Mouse) Tissue Lysate with Human beta III Tubulin peptide (ab18660) at 2 µg/ml
    Lane 6 : Brain (Rat) Tissue Lysate with Human beta III Tubulin peptide (ab18660) at 2 µg/ml

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds
  • All lanes : Anti-beta III Tubulin antibody (ab18207) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 55 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab18207 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody , and visualised using ECL development solution ab133406

  • ab18207 staining beta III Tubulin in Neuro-2a cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18207 at 1μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
    Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Immunohistochemistical staining (Formaldehyde/PFA-fixed paraffin-embedded sections) for Neuron specific beta III Tubulin antibody - Neuronal Marker (ab18207) on Dogfish/Catshark Tissue sections (head: snout region). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody used at 1/2000 incubated for 2 hours at RT. Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).
  • ab18207 staining beta III Tubulin in SKNSH cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18207 at 1μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
    Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Overlay histogram showing U-87MG cells stained with ab18207 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18207, 0.01μg/1x106) for 30 min at 22ºC. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.01μg/1x10cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in U-87MG cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • ab18207 at 1/2000 staining rat cerebellum tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed prior to incubation with the antibody for 16 hours. A biotinylated goat polyclonal antibody was used as the secondary.

    See Abreview

  • ab18207 staining beta III tubulin in PC-12 cells treated with venlafaxine hydrochloride (ab120715), by ICC/IF. Increase in the number and length of neurites (stained with beta III tubulin) correlates with increased concentration of venlafaxine hydrochloride, as described in literature.
    The NGF treated cells were incubated at 37°C for 6 hour in media containing different concentrations of ab120715 (venlafaxine hydrochloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18207 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899) secondary antibody at 1/250 dilution was used.

  • All lanes : Anti-beta III Tubulin antibody (ab18207) at 1/1000 dilution

    All lanes : Mouse hippocampus tissue lysate

    Lysates/proteins at 8 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG (H&L) at 1/5000 dilution

    Predicted band size: 50 kDa
    Observed band size: 55 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds

    See Abreview

  • ab18207 at 1/2000 staining mouse brain tissue sections by IHC-P. The tissue was formaldehyde fixed and an enzymatic antigen retrieval step was performed prior to incubation with the antibody for 16 hours. A biotinylated goat anti-rabbit IgG was used as the secondary.

    See Abreview

参考文献

This product has been referenced in:

See all 228 Publications for this product

レビューと Q&A

1-5 of 5 Q&A

Question
Answer



We have never used the peptide ab18660 to block ab117740, and therefore I cannot guarantee it will be valid for this purpose.

However, the peptide ab18660 was used to raise antibody ab18207, so it could be used as a blocking peptide for it.

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Answer

Thank you for contacting Abcam.

I've compiled a list of primary antibodies that have been tested in mouse tissue via western blot. The only ovary-specific antibodies that I found were anti-human (with only 50% homology with the respective mouse protein), so the ovary antibody listed below is not exclusively-specific to ovary but also gives a positive signal in kidney, placenta, and small intestine:

1. Liver: anti-albumin ab135575 (https://www.abcam.com/albumin-antibody-c-terminal-ab135575.html)
2. Brain: anti-neuron specific beta III tubulin ab18207 (https://www.abcam.com/neuron-specific-beta-iii-tubulin-antibody-neuronal-marker-ab18207.html)
3. Kidney: anti-SLC14A2 ab95365 (https://www.abcam.com/slc14a2-antibody-ab95365.html)
4. Testis: anti-histone H2B ab23913 (https://www.abcam.com/histone-h2b-testis-specific-antibody-ab23913.html)
5. Ovary: anti-ENPP2 (also kidney, placenta, small intestine): ab77104 (https://www.abcam.com/enpp2-antibody-5h3-ab77104.html)

The advantage of this combination of antibodies is that you can use the same anti-rabbit secondary since they were all raised in a rabbit host. Our anti-rabbit secondary antibodies are listed in the link below:
https://www.abcam.com/Search?Keywords=anti-rabbit&fReset=1&pt=-1&source=TopSearch

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link:
https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

You may be able to reduce your background staining by blocking with 10% normal serum from the host species of your secondary antibody for 1 hour. For example, if you were using a goat anti-rabbit secondary antibody, I would recommend using normal goat serum (ab7481) for blocking.

What was the dilution of the primary antibody? It can often also help to try using less antibody if you would like to reduce the background staining. Was your no primary antibody control clean? What species were your cells from and how long were they fixed?

I hope this helps, if not, I will be happy to help you further once I have the above information.

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Question

Sehr geehrte Damen und Herren,

ich habe einige Anfragen zu folgenden 1. Antikörpern

1)
ab31232 Anti-Glutamate Receptor 1 (AMPA subtype)
ab18207 Anti-Neuron specific beta III Tubulin antibody- Neuronal Marker

Diese Antikörper sollen innerhalb von 2 Wochen aliquotiert werden und bei -20°C oder -80°C° gelagert werden. Könnten wir diese Antikörper auch mit PBS (z.B. 0,1 m) aliquotieren, zum Beispiel 1:1, das würde uns bezüglich des Verbrauchs entgegen kommen.
Wie häufig kann man das Antikörper Aliquots auftauen. Wo liegt Ihre Empfehlung, da man es so wenig wie möglich machen soll.

2)
ab8882 Serotonin (conjgated) antbody
ab5836 Histamine (conjugated) anitbody

Im Produkt sheet steht unter "storage buffer" Constituents 50% Glycerol, ddH2O
Wie ist das zu verstehen, heißt es das 50% der Menge an Antikörper Glycerol (Glycerin) enthalten, so dass wir z. Beispiel 2:1000, statt 1:1000 beim Antikörperansatz verwenden müßten.

Könnten diese Antkörper auch aliquotiert und eingefroren werden, wenn ja wie.

Über eine schnelle Beantwortung würde ich mich sehr freuen.

Mit freundlichen Grüßen

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2011-11-10T07:00:00Z
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MicrosoftInternetExplorer4




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Answer

Vielen Dank für Ihre Anfrage.

1.) ich kann generell nicht empfehlen, Antikörper zu verdünnen bevor sie eingefroren werden. Eine geeignete Proteinkonzentration ist wichtig für die Stabilität der Antikörper und wird deutlich verringert wenn der Antikörper verdünnt wird.

Ich stimme Ihnen zu, dass je weniger "freeze/thaw" Zyklen die Antikörper mitmachen müssen desto besser. Genaue Zahlen kann ich Ihnen leider nicht geben, aber meine persönliche Erfahrung spricht für weniger als 5 bei manchen Antikörpern während andere sich als ausnehmend stabil erweisen.

2.) Glycerol wird verwendet um Antikörperlösungen bei _20 oder -80C lagern zu könne ohne das die Lösung einfriert/durchfriert. Deshalb brauchen diese Antiköper auch nicht aliquotiert zu werden.

Leider müssen Arbeitskonzentrationen immer optimiert werden. Aber grundsätzlich muss von diesen Antikörpern nicht mehr verwendet werden, da die Glycerolmenge in der Konzentration des Stocks berücksichtigt ist.

Ich hoffe, diese Information ist hilfreich und wünsche Ihnen viel Erfolg bei Ihren versuchen.

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Answer

I am sorry to hear that the glass jars did crack, these usually handle the heating well.

An extended incubation may work however I have not tested this condition and cannot guarantee success with this condition. If you have other means of submersing the slides, such as plastic staining trays, you may try to use those to perform the antigen retrieval in the microwave.

Please let me know if you have any further questions or concerns.

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