Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
重组RabMAb

重组Anti-SLP76 (phospho Y145)抗体[EP2853Y] (ab75829)

Reviews (1)Q&A (3)References (8)
Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Dot Blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Flow Cytometry (Intracellular) - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
  • Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2853Y] to SLP76 (phospho Y145)
  • Suitable for: ICC/IF, Dot blot, WB, Flow Cyt (Intra)
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-SLP76 (phospho Y145)抗体[EP2853Y]
    参阅全部 SLP76 一抗

  • 描述

    兔单克隆抗体[EP2853Y] to SLP76 (phospho Y145)

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, Dot blot, WB, Flow Cyt (Intra)more details
    不适用于: IHC-P

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • Jurkat whole cell lysate (ab7899) treated with pervanadate.

  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • 存储溶液

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    EP2853Y

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab75829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
ICC/IF
1/500.
Dot blot
1/1000.
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
说明
ICC/IF
1/500.
Dot blot
1/1000.
WB
1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Involved in T-cell antigen receptor mediated signaling.

  • 组织特异性

    Highly expressed in spleen, thymus and peripheral blood leukocytes. Highly expressed also in T-cell and monocytic cell lines, expressed at lower level in B-cell lines. Not detected in fibroblast or neuroblastoma cell lines.

  • 序列相似性

    Contains 1 SAM (sterile alpha motif) domain.
    Contains 1 SH2 domain.

  • 结构域

    The SH2 domain mediates interaction with SHB.

  • 翻译后修饰

    Phosphorylated after T-cell receptor activation by ZAP-70.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 76 kDa tyrosine phosphoprotein antibody
      • CG8697 antibody
      • LCP 2 antibody
      • LCP2 antibody
      • LCP2_HUMAN antibody
      • Lymphocyte cytosolic protein 2 antibody
      • SH2 domain containing leukocyte protein 76 KD antibody
      • SH2 domain containing leukocyte protein of 76kD antibody
      • SH2 domain containing leukocyte protein of 76kDa antibody
      • SH2 domain-containing leukocyte protein of 76 kDa antibody
      • SLP 76 antibody
      • SLP 76 tyrosine phosphoprotein antibody
      • SLP-76 antibody
      • SLP-76 tyrosine phosphoprotein antibody
      • SLP76 antibody
      • SLP76 tyrosine phosphoprotein antibody
      see all

    图片

    • Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/5000 dilution

      Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
      Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. treated with pervanadate at 50nM for 5 min.
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. Treated with pervanadate at 50nM for 5 min. Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 60 kDa
      Observed band size: 76 kDa why is the actual band size different from the predicted?



      Blocking buffer 5% NFDM/TBST. Diluting buffer 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

      ab75829 staining SLP76 in Jurkat by Immunocytochemistry. Tissue was fixed with 4% paraformaldehyde. Samples were incubated with primary antibody (4.5 μg/ml). ab150077 AlexaFluor®488 Goat anti-Rabbit (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 2.5 μg/ml and DAPI were used as counter stains.

      Confocal image showing the expression was increased after treatment with pervanadate (1mM, 30min) and then decreased after treatment with the Lambda Protein Phosphatase 31℃ for 2h

    • Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

      ab75829 staining SLP76 (phospho Y145) antibody in both treated (50mM of pervanadate for 5 minutes) and untreated Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000.

      DAPI was used as a nuclear counterstain.

    • Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/200000 dilution

      Lane 1 : Jurkat cell lysate un-treated
      Lane 2 : Jurkat cell lysate treated with pervanadate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 60 kDa
      Observed band size: 76 kDa why is the actual band size different from the predicted?

    • Dot Blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Dot Blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

      Dot blot analysis of SLP76 (phospho Y145) phospho peptide (Lane 1), SLP76 Non-phospho peptide (Lane 2), labelling SLP76 (phospho Y145) with ab75829 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and diluting buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Flow Cytometry (Intracellular) - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Flow Cytometry (Intracellular) - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

      Intracellular flow cytometric analysis of permeabilized Jurkat cells, untreated (green) or pervanadate treated (red) using ab75829 at a 1/100 dilution. Also, pervanadate Jurkat cells using the same antibody preincubated with phospho SLP76 peptide (blue) or non phospho SLP76 peptide (orange).

       

    • Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)
      Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

    文献 (8)

    发表研究结果有使用 ab75829？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab75829 被引用在 8 文献中.

    • Sadhu L  et al. ARPC5 isoforms and their regulation by calcium-calmodulin-N-WASP drive distinct Arp2/3-dependent actin remodeling events in CD4 T cells. Elife 12:N/A (2023). PubMed: 37162507
    • Rossatti P  et al. Rapid increase in transferrin receptor recycling promotes adhesion during T cell activation. BMC Biol 20:189 (2022). PubMed: 36002835
    • Zeng L  et al. PLCγ1 promotes phase separation of T cell signaling components. J Cell Biol 220:N/A (2021). PubMed: 33929486
    • Zhou J  et al. Identification and Characterization of the Amphioxus Lck and Its Associated Tyrosine Phosphorylation-Dependent Inhibitory LRR Receptor. Front Immunol 12:656366 (2021). PubMed: 34149695
    • Dong R  et al. Rewired signaling network in T cells expressing the chimeric antigen receptor (CAR). EMBO J 39:e104730 (2020). PubMed: 32643825
    • Wang QL  et al. T Cell Receptor (TCR)-Induced PLC-?1 Sumoylation via PIASxß and PIAS3 SUMO E3 Ligases Regulates the Microcluster Assembly and Physiological Function of PLC-?1. Front Immunol 10:314 (2019). PubMed: 30873169
    • Yang CY  et al. Dual-specificity phosphatase 14 (DUSP14/MKP6) negatively regulates TCR signaling by inhibiting TAB1 activation. J Immunol 192:1547-57 (2014). PubMed: 24403530
    • Gill T & Levine AD Mitochondria-derived hydrogen peroxide selectively enhances T cell receptor-initiated signal transduction. J Biol Chem 288:26246-55 (2013). PubMed: 23880762

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-SLP76 (phospho Y145) antibody [EP2853Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    100000 cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - whole cell (jurkats)
    Specification
    jurkats
    Treatment
    TCR activation with OKT3 and anti CD8
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°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.

    Dr. Tejpal Gill

    Verified customer

    提交于 May 13 2014

    Question

    The customer wishes to get a refund for the antibody and claims she has tried several dilutions within the range you recommended but still wasn't able to get good results.

    Please advise.
    Regards,

    Read More
    Answer

    Thank you for contacting us.

    Your credit note ID is 23976.

    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Question

    Here is the customer reply as attached files.
    Thank for your continuous help.


    1) Abcam product code ab75829

    2) Abcam order reference number: 6740/12

    3) Description of the problem: bands for p-SLP76 are not seen but only not specific bands

    4) Sample preparation:

    Type of sample (whole cell lysates, fraction, recombinant protein…): jurkat cell lysates.

    Lysis buffer: Tris HCl 50mM pH=7.5, NaCl 150mM, MgCl2 5mM, EDTA 2mM pH=7.5, triton 1%

    Protease inhibitors: protease inhibitor cocktail (P8340 sigma)

    Phosphatase inhibitors: phosphatase inhibitor cocktail (P5726 sigma)

    Reducing agent

    Boiling for ≥5 min? 5min

    Protein loaded 10ug/lane and 20ug/lane

    Positive control ---

    Negative control daudi and raji cell lysates

    5) Percentage of gel: lower- 10%, upper- 4%

    Type of membrane: cellulose nitrate membrane 0.20µm, 69X82mm

    Protein transfer verified

    Blocking agent and concentration: difco skim milk 5% in TBSX1

    Blocking time: 1hour

    Blocking temperature: room tem

    6) Primary antibody (If more than one was used, describe in “additional notes”): anti-SLP76 (phospho Y145) antibody EP2853Y

    Concentration or dilution: 1:500 in TBSTX1 and 5% BSA, 1:1000 in TBSTX1 and 5% BSA

    Diluent buffer

    Incubation time: over night

    Incubation temperature: 4Cº



    7) Secondary antibody: goat anti-rabbit IgG (H+L)-HRP conjugate

    Species:

    Reacts against:

    Concentration or dilution: 1:4000 in TBSX1

    Diluent buffer

    Incubation time: 1-1.5 hour

    Incubation temperature: room tem

    Fluorochrome or enzyme conjugate:



    8) Washing after primary and secondary antibodies:

    Buffer: TBSTX1

    Number of washes: 3 washes each one 7 min



    9) Detection method: EZ-ECL chemiluminescence detection kit for HRP



    10) How many times have you run this staining? twice

    Do you obtain the same results every time? yes

    What steps have you altered to try and optimize the use of this antibody? I used different concentration of the antibody and protein loading.

    Regards,

    Read More
    Answer

    Thank you for your enquiry regarding ab75829 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 have discussed this case with the Lab. The protocol looks absolutely fine, the only recommendation we could make to dilute the antibody less if it is possible: 1/1000- 10,000.

    I am sorry for this inconvenience!

    Should your customer still has any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

    Read More

    Question

    Dear Tech support

    One of our customers has ordered ab75829 but hasn't received a band with the recommended dilutions of 1/100000 or 1/200000.
    These dilutions are considered very high, could it be an error? In your opinion, after trying with these dilutions, what dilutions should the customer try next?

    Please advise.
    Thank you for you help.
    Regards,

    Read More
    Answer

    Thank you for your inquiry.

    The dilution factors for each application listed on the product datasheet for ab75829 are what we currently recommended, it is still relevant.

    If you wish me take this case further, 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.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com