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重组RabMAb

重组Anti-BRN3A抗体[EP1972Y] (ab81213)

Reviews (6)Q&A (1)References (17)
Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
  • Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)
  • Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
  • Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
  • Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)
  • Anti-BRN3A antibody [EP1972Y] (ab81213)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1972Y] to BRN3A
  • Suitable for: Flow Cyt (Intra), WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-BRN3A抗体[EP1972Y]
    参阅全部 BRN3A 一抗

  • 描述

    兔单克隆抗体[EP1972Y] to BRN3A

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WBmore details
    不适用于: ICC/IF or IP

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human BRN3A aa 300-400 (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: Human brain lysate; SW480; Mouse cerebrellum; Rat brain; Rat spinal cord. ICC/IF: HeLa cells. Flow Cyt (intra): SH-SY5Y cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab81213于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/90.
WB (1)
1/2000. Predicted molecular weight: 43 kDa.

For unpurified use at 1/500 - 1/1000. 
说明
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/90.
WB
1/2000. Predicted molecular weight: 43 kDa.

For unpurified use at 1/500 - 1/1000. 
应用说明
Is unsuitable for ICC/IF or IP.

靶标

  • 功能

    Probable transcription factor which may play a role in the regulation of specific gene expression within a subset of neuronal lineages. May play a role in determining or maintaining the identities of a small subset of visual system neurons.

  • 组织特异性

    Brain. Seems to be specific to the retina. Present in the developing brain, spinal cord and eye.

  • 序列相似性

    Belongs to the POU transcription factor family. Class-4 subfamily.
    Contains 1 homeobox DNA-binding domain.
    Contains 1 POU-specific domain.

  • 发展阶段

    Expression peaks early in embryogenesis (day 13.5) and is undetectable 14 days after birth.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • Brain specific homeobox/POU domain protein 3A antibody
    • Brain-3A antibody
    • Brain-specific homeobox/POU domain protein 3A antibody
    • Brn 3.0 antibody
    • BRN 3A antibody
    • Brn-3A antibody
    • Brn3 antibody
    • BRN3A antibody
    • class 4 antibody
    • FLJ13449 antibody
    • Homeobox/POU domain protein RDC 1 antibody
    • Homeobox/POU domain protein RDC-1 antibody
    • Homeobox/POU domain protein RDC1 antibody
    • Oct T1 antibody
    • Oct-T1 antibody
    • OctT1 antibody
    • PO4F1_HUMAN antibody
    • POU class 4 homeobox 1 antibody
    • POU domain antibody
    • POU Domain Class 4 Transcription Factor 1 antibody
    • Pou4f1 antibody
    • RDC 1 antibody
    • RDC1 antibody
    • transcription factor 1 antibody
    see all

图片

  • Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
    Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
    All lanes : Anti-BRN3A antibody [EP1972Y] (ab81213) at 1/2000 dilution (purified)

    Lane 1 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse cerebellum lysates
    Lane 3 : Rat brain lysates
    Lane 4 : Rat spinal cord lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)
    Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)

    Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling BRN3A with purified ab81213 at 1/100 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
    Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
    All lanes : Anti-BRN3A antibody [EP1972Y] (ab81213) (unpurified)

    Lane 1 : LNCaP
    Lane 2 : SW480
    Lane 3 : PC-3
    Lane 4 : Human brain lysate

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

  • Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)
    Western blot - Anti-BRN3A antibody [EP1972Y] (ab81213)This image is courtesy of an Anonymous Abreview.
    All lanes : Anti-BRN3A antibody [EP1972Y] (ab81213) at 1/2000 dilution (unpurified)

    Lane 1 : Adult mouse cerebellum lysate
    Lane 2 : Adult mouse lower brainstem lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated Mouse anti-Rabbit IgG monoclonal. at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa


    Exposure time: 2 minutes

    See Abreview

  • Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)
    Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EP1972Y] (ab81213)

    Overlay histogram showing SH-SY5Y cells stained withunpurified ab81213 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab81213, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. 

     

     

  • Anti-BRN3A antibody [EP1972Y] (ab81213)
    Anti-BRN3A antibody [EP1972Y] (ab81213)

文献 (17)

发表研究结果有使用 ab81213？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab81213 被引用在 17 文献中.

  • Zhao Q  et al. Identification and verification of a prognostic signature based on a miRNA-mRNA interaction pattern in colon adenocarcinoma. Front Cell Dev Biol 11:1161667 (2023). PubMed: 37745305
  • Lindborg JA  et al. Optic nerve regeneration screen identifies multiple genes restricting adult neural repair. Cell Rep 34:108777 (2021). PubMed: 33657370
  • Zhang M  et al. Brain and Retinal Abnormalities in the 5xFAD Mouse Model of Alzheimer's Disease at Early Stages. Front Neurosci 15:681831 (2021). PubMed: 34366774
  • Bataille A  et al. In Vitro Differentiation of Human Skin-Derived Cells into Functional Sensory Neurons-Like. Cells 9:N/A (2020). PubMed: 32316463
  • Mak HK  et al. MicroRNA-19a-PTEN Axis Is Involved in the Developmental Decline of Axon Regenerative Capacity in Retinal Ganglion Cells. Mol Ther Nucleic Acids 21:251-263 (2020). PubMed: 32599451
View all Publications for this product

客户评价及客户问答

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1-7 of 7 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BRN3A antibody [EP1972Y]

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Retina)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citrate buffer
Permeabilization
Yes - tween 20
Specification
Retina
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Davidson
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 05 2018

Immunohistochemistry (Frozen sections) abreview for Anti-BRN3A antibody [EP1972Y]

 Poor
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse eye)
Permeabilization
Yes - 0.3% triton X-100
Specification
mouse eye
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Mar 18 2016

Immunohistochemistry (Frozen sections) abreview for Anti-BRN3A antibody [EP1972Y]

 Poor
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Retinal ganglion cells)
Permeabilization
Yes - Triton-X-100
Specification
Retinal ganglion cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. XXXXXX XXXXXX

Verified customer

提交于 Aug 11 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-BRN3A antibody [EP1972Y]

 Excellent
Abreviews
This product is known to not work in this application or species.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Dorsal Root Ganglia (DRG) neurons)
Specification
Dorsal Root Ganglia (DRG) neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - tween 20
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 18 2012

IHC - Wholemount abreview for Anti-BRN3A antibody [EP1972Y]

 Good
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (Brain explant)
Specification
Brain explant
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 05 2012

Western blot abreview for Anti-BRN3A antibody [EP1972Y]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (cerebellum & lower portion of brainstem (adult))
Loading amount
20 µg
Specification
cerebellum & lower portion of brainstem (adult)
Gel Running Conditions
Reduced Denaturing (12 %)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 15 2011

Question

We used 5% goat blocking serum, and preincubated the sections for 30min, and the wholemount for 2 days. The secondaries have both been used successfully with other primaries over the past 2-3 months. The protocol for the sections is below:



Slide 1 – Abcam rb α Brn3a 1:100; g α rb Alexa 594 1:400



Slide 2 – Abcam rb α Brn3a 1:100; g α rb Alexa 488 1:400



· Let frozen slides air dry for 10min, circle with PAP pen

· Label slides with pencil (primary and secondary antibody, concentrations and date)


Wash 3x in PBS (5 min each), coplin jars

Triton X-100 3% (15 min)

Wash 3x in PBS (5 min each), coplin jars

Pre-incubate in 120 µl of 5% Goat Blocking Serum and cover with parafilm for 30 min at RT

Prepare primary antibodies by adding primary AB to NEW blocking buffer (total volume 100 µl) For double label: need to halve concentration denominator since applying both antibodies together so there is double the volume. Make as a batch for each antibody. Vortex all tubes. Mix double label tubes before applying.




rb α Brn3a, 1:1 → 1:100 (2/6/2013, Abcam #AB81213)

2 µl 1:1 + 198 µl GBS

Gives 200 µl 1:100 total, use 90 µl for each slide: Slide 1, Slide 2




Pipette 90 µl of the appropriate prepared primary AB onto slides, cover in parafilm for 2 h at RT



Wash 3x in PBS (5 min each), coplin jars

Make up secondary antibodies by adding secondary AB to NEW blocking buffer (total volume 100 µl). Use Alexa 594 or 488-coupled antibody at 1:400.




g α rb Alexa 594 1:1 → 1:400

2 µl undiluted + 798 µl GBS

Gives 100 µl 1:400 for Slide 1



g α rb Alexa 488 1:1 → 1:400

2 µl undiluted + 798 µl GBS

Gives 100 µl 1:400 for Slide 2




Centrifuge antibodies at 12000 G for 5 min

Incubate sections in secondary antibodies (90 µl) for 1 hour at RT covered with parafilm, keep in dark

Wash 3x in PBS (15 min each) in dark

Coverslip on slide with drop of vectashield with DAPI. Keep in dark.

Read More
Answer

With the frozen sections, did you do a fix and permeabilize step. I didnt notice that in your protocol. If you haven't then that 's probably the reason for no staining. Its important to do that so that the epitope is accessible.
I would also just incubate the primary antibody at 4 degrees overnight.

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