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Immunology Immunoglobulins Heavy Chain IgG

山羊抗兔IgG H&L (Alkaline Phosphatase) (ab6722)

  • Datasheet
  • SDS
Submit a review Q&A (6)References (40)

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Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
  • ELISA - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
  • Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
  • Dot Blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
  • Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

Key features and details

  • Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase)
  • Conjugation: Alkaline Phosphatase
  • Host species: Goat
  • Isotype: IgG
  • Suitable for: IHC-P, WB, ELISA, Dot blot, ICC/IF, IHC-Fr, ICC

Conjugates logo Related conjugates and formulations

Agarose Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 680 Alexa Fluor® 790 APC beta-galactosidase Biotin Cy2 ® Cy3 ® Cy3.5 ® Cy5 ® Cy5.5 ® DyLight® 488 DyLight® 550 DyLight® 594 DyLight® 650 FITC Glucose Oxidase Gold 12nm Gold 6nm HRP polymer IRDye® 680RD IRDye® 800CW PE Texas Red ® TRITC Unconjugated Unconjugated Unconjugated Unconjugated Unconjugated

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概述

  • 产品名称

    山羊抗兔IgG H&L (Alkaline Phosphatase)
    参阅全部 IgG 二抗
  • 宿主

    Goat
  • 靶标种属

    Rabbit
  • 经测试应用

    适用于: IHC-P, WB, ELISA, Dot blot, ICC/IF, IHC-Fr, ICCmore details
  • 免疫原

    Rabbit IgG whole molecule

  • 偶联物

    Alkaline Phosphatase

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C.
  • 存储溶液

    Preservative: 0.1% Sodium azide
    Constituents: 0.00136% Zinc chloride, 0.0095% Magnesium chloride, 0.79% Tris HCl, 50% Glycerol, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...
  • 纯度

    Affinity purified
  • 纯化说明

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • 共轭说明

    Alkaline Phosphatase (Calf Intestine) (Molecular Weight 140,000 daltons)
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Secondary antibodies
    • anti-Rabbit
    • IgG
    • Enzyme
    • Alkaline Phos

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab6722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
IHC-P
Use at an assay dependent dilution.
WB
1/1000 - 1/4000.
ELISA
1/3000 - 1/5000.
Dot blot
Use at an assay dependent dilution.
ICC/IF
1/200 - 1/1000.
IHC-Fr
Use at an assay dependent dilution.
ICC
Use at an assay dependent dilution.
说明
IHC-P
Use at an assay dependent dilution.
WB
1/1000 - 1/4000.
ELISA
1/3000 - 1/5000.
Dot blot
Use at an assay dependent dilution.
ICC/IF
1/200 - 1/1000.
IHC-Fr
Use at an assay dependent dilution.
ICC
Use at an assay dependent dilution.

图片

  • Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
    Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

    ab6722 was used at dilution 1/10000 with the primary antibody ab16731 in WB. See the review on ab16731.

  • ELISA - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
    ELISA - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

    ab6722 was used with the primary antibody ab2594 in ELISA. See Abreview on ab2594.

  • Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
    Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

    Western Blot of Anti-Rabbit IgG (H&L) (GOAT) Antibody.

    Lane M: 3 µl Molecular ladder.

    Lane 1: Rabbit IgG whole molecule.

    Lane 2: Rabbit IgG F(ab) Fragment.

    Lane 3: Rabbit IgG F(c) Fragment.

    Lane 4: Rabbit IgM Whole Molecule.

    Lane 5: Normal Rabbit Serum.

    All samples were reduced. Load: 50 ng per lane. Blocked for 30 min at RT.

    Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40,000 for 30 min at RT.

    Predicted/Obsevered Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM.

    Rabbit F(c) migrates slightly higher.

  • Dot Blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
    Dot Blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

    Dot Blot of ab6722.

    Antigen: Rabbit IgG.

    Load: Lane 1 - 200ng; Lane 2 - 66.7ng; Lane 3 - 22.2ng; Lane 4 - 7.4ng; Lane 5 - 2.5ng.

    Secondary antibody: ab6722 used at 1:1,000 for 60 min at RT. Blocked for 60 min at RT.

    Reaction visualized using alkaline phosphatase substrate for 30 seconds at RT.

  • Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
    Western blot - Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)

    Lane 1: Rabbit IgG.

    Load: 100 ng per lane.

    Secondary antibody: ab6722 used at 1:1,000 for 60 min at RT. Blocked for 30 min at RT.

    Predicted/Observed size: 55 and 28 kDa, 55 kDa for Rabbit IgG.

实验方案

  • ELISA procedure for ab6722

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (40)

发表研究结果有使用 ab6722?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab6722 被引用在 40 文献中.

  • Wawrzykowski J  et al. The comparison of pro- and antioxidative parameters in plasma and placental tissues during early phase of placental development in cows. Mol Biol Rep 48:1291-1297 (2021). PubMed: 33507474
  • Hao J  et al. Curcumin suppresses colorectal tumorigenesis via the Wnt/ß-catenin signaling pathway by downregulating Axin2. Oncol Lett 21:186 (2021). PubMed: 33574925
  • Palomino J  et al. Embryo Buoyancy and Cell Death Gene Expression During Embryogenesis of Yellow-Tail Kingfish Seriola lalandi. Front Cell Dev Biol 9:630947 (2021). PubMed: 33816479
  • Lee JS  et al. A Comparison of Isolation Stress and Unpredictable Chronic Mild Stress for the Establishment of Mouse Models of Depressive Disorder. Front Behav Neurosci 14:616389 (2020). PubMed: 33488368
  • Gritsyna YV  et al. Inhibition of Histone Deacetylases 4 and 5 Reduces Titin Proteolysis and Prevents Reduction of TTN Gene Expression in Atrophied Rat Soleus Muscle after Seven-Day Hindlimb Unloading. Dokl Biochem Biophys 495:338-341 (2020). PubMed: 33368047
View all Publications for this product

客户评价及客户问答

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1-6 of 6 Abreviews or Q&A

Question

Do I need to add a stop solution (ie NaOH) at the end of the sandwich ELISA after the colour develops with ALP chromogen or that is not necessary?

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Abcam community

Verified customer

Asked on Feb 03 2012

Answer

Thank you for contacting us.

Just as with a common indirect ELISA it is important to stop the reaction using the appropriate stop solution.

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

Read More

Abcam Scientific Support

回复于 Feb 03 2012

Question

I repeated the experiment and diluted plasma samples out to 1:4800 fold but this dilution dependent signal increase phenomenon persisted. In other words, the signal from the 1:4800 fold dilution was higher than the 1:2 dilution! I really doubt if this is a hook effect as I would expect the signal to decrease again after adequate dilution and not continue to rise. Then I thought whether it could be non-specific binding of detection ab (I used ab6722 - a Goat anti-rabbit ab because my primary is Rabbit anti-human). So I included controls in which I added only the secondary ab to plasma-coated wells, without any primary. I saw moderate signal from these wells (ie higher than BSA negative controls but weaker than corresponding plasma-coated wells with both primary and secondary added). Does this mean the secondary ab is binding non-specifically to something in the human plasma? Are you aware of cross-reacitivity of this Goat anti-rabbit ab (ab6722) with any human Ig??

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Abcam community

Verified customer

Asked on Mar 23 2012

Answer

I am not aware of any cross reactivity of this antibody with human Ig. However we do offer a number of secondary antibodies which are designed to reduce background levels by either being pre-absorbed against other proteins and Igs or by digestion into F(ab) and Fc fragments. I've included links to a few of those below.

That said I do not feel that the background you are experiencing should be the cause for the type of response you have been seeing. This is usually something that can be easily subtracted from your results if you run 2 or 3 wells with this control as a baseline.

Another possibility is that it could be that the dilution buffer you are using is somehow causing this reaction. What buffer are you using? Have you tried using filtering this or using another buffer?

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

These are links to the antibodies that have been pre-absorbed:

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-Alkaline-Phosphatase-pre-adsorbed-ab97072.html

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-Alkaline-Phosphatase-pre-adsorbed-ab97072.html

https://www.abcam.com/Goat-F-ab-2-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-AP-pre-adsorbed-ab98469.html

Read More

Abcam Scientific Support

回复于 Mar 23 2012

Question

Many thanks for your reply. I really appreciate your suggestions. My detection is working well now. However I am facing some strange results which I would love your thoughts on please:   I am struggling to interpret an indirect ELISA I am making. It involved coating wells with human plasma overnight, blocked with milk the next day, then primary ab (Abcam), then detection ab (Abcam) and finally the ALP substrate. I ran parallel wells in which I coated with   1)      known concentration of antigen which is a recombinant protein to generate my standard curve and 2)      BSA (negative control) I got nice reproducible standard curves (R2=0.99) and uniformly negative signals from BSA. So all good. However I am seeing a dose-dependent increase in signal in plasma samples. As plasma dilution increased, signal increased - in other words, the more dilute the plasma (so in theory lesser amount of antigen) - the stronger the signal! I could not understand why. I repeated the experiments and I am seeing it every time (4x now). At first I thought it could be due to primary ab binding to "unoccupied sites in the wells" directly, which then generated a signal when secondary added, which would explain why the signal is stronger in diluted sample. But if that's the case I should see that in the BSA wells, but I am not. I did block the wells with milk after coating to ensure all unbound sites are occupied. Then I thought could it be something in the plasma which is inhibiting the substrate reaction (ALP). So the more dilute the plasma, the less the inhibition, and the stronger the signal. But plasma was only used to coat the plate overnight. The plates were then washed after every step with TBS-tween (2-4 times) after every step. So I would imagine not much of any “inhibitors” are around by the time substrate is added. Then I thought whether it could be a “hook” effect. In other words, the plasma is too concentrated and antigen are crowding up thus reducing the amount of binding. But I thought usually plasma then need to be diluted up to 100 folds to relieve the hook. But I am seeing a “nice” dose dependent increase in signal as plasma was diluted from 1:2, 1:4, 1:8 to 1:16 only.

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Abcam community

Verified customer

Asked on Mar 21 2012

Answer

Thank you for contacting me again. It is always a pleasure to hear from you.

After reviewing your email, I am lead to believe that this really is a hook effect as you had mentioned. I am not an expert on this hook effect in plasma specifically, and so cannot comment on the 100 fold dilutions usually required, but your description does sound like a hook effect. These should eventually tail off with further dilutions.

The hook effect may also be created by overcoating with primary antibody; you may wish to further dilute your primary to tackle this as well.

I hope these suggestion help. Let me know if you have any questions.

Read More

Abcam Scientific Support

回复于 Mar 21 2012

Question

Thanks for your advice and suggestions last time. I got another ab7468 and it is working well, both for WB and my reattempt ELISA.   However I have a question which I hope you may have an answer to:   After adding the pNPP substrate, I waited for 30 min, as recommended, but the signal was very weak. OD was around 0.1 and negative control 0.06. However as I waited for longer, up to 1.5 hour, the signal intensified in the samples and went up to about 0.4. Negative control still remained low at 0.06, which was good. The resultant standard curve was great (R2=0.99). However I could not understand why it took almost 60-90 min before the OD got up to a “good signal”. I don’t think I am simply increasing background as the standard curve was good and the longer duration did not intensify signal from any of the negative controls.   Is this longer than usual duration due to 1)      Inadequate primary ab? 2)      Inadequate secondary ab? 3)      Inadequate coating antigen on plate??  

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Abcam community

Verified customer

Asked on Mar 14 2012

Answer

Thank you for contacting us, I am glad to hear that the new product is working well for you.

Usually when color is developing slowly it is one of three issues. 1) the plates and reagents have not come to room temperature. 2) the conjugates are weak, try to prepare these right before use and at correct concentration 3) the presence of peroxidases, azide or other contaminants are interfering with the reaction, these may be removed by dyalisis, gel filtration or with purification columns.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More

Abcam Scientific Support

回复于 Mar 14 2012

Question

I recently purchased your ab (ie Rabbit anti-human FNDC5 IgG), to be used in a ELISA. I am wondering what detection system would you recommend?   Ie According to the Abcam ELISA protocol, one option is HRP.   So Do I add a secondary ab conjugated (ie anti-rabbit IgG Fc, HRP conjugated), followed by washing and then dispensing 100 ul of substrate solution (ie TMB?)   Or would you recommend other secondary/substrate solution system?  

Read More

Abcam community

Verified customer

Asked on Feb 02 2012

Answer

Thank you for contacting Abcam.

Enzymatic detection with an antibody that has been conjugated to HRP or to AP is a widely used and robust method. As this target is located on the peroxisome membrane I would recommend using anAlkaline Phosphataseconjugated secondary for this target prehaps ab6722,Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (AP). Followed by an appropriate substrate detection of your choice. I recommend theAlkaline Phosphatase chromogen (BCIP/NBT). We do offer this as a ready to use product, ab7468.

I hope you have found this information helpful. Please let me know if you have any questions. I have places links to each of the products I have mentioned here below.

Ab6722:https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-AP-ab6722.html

Ab7468:https://www.abcam.com/Alkaline-Phosp

Read More

Abcam Scientific Support

回复于 Feb 02 2012

Question

I would like to receive a vial of ab34767 as a refund for my order for ab28664. Also, this is my purchasing code for another antibody I have ordered and that does not work with the replacement product you will send me :Po-IG/101857, order on the 18th August 2011 and for which I would like a refund or credits.   Please advise me how quickly the replacement antibody will arrive, as I would like to proceed with my experiments.   Thank you for your assistance!

Read More

Abcam community

Verified customer

Asked on Sep 30 2011

Answer

Thank you for your reply. I have sent a free vial of ab34767 to replace ab28664. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. 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 have also issued a credit note for ab6722. 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

Abcam Scientific Support

回复于 Sep 30 2011

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