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  1. Link

    products/secondary-antibodies/goat-chicken-igy-hl-hrp-ab97135.pdf

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Secondary antibodies anti-Chicken IgY Enzyme HRP
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Goat Anti-Chicken IgY H&L (HRP) (ab97135)

  • Datasheet
  • SDS
Reviews (2)Q&A (4)References (24)

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Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
  • Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
  • Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)

Key features and details

  • Goat Anti-Chicken IgY H&L (HRP)
  • Conjugation: HRP
  • Host species: Goat
  • Isotype: IgG
  • Suitable for: ICC, IHC-P, ELISA, WB

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Overview

  • Product name

    Goat Anti-Chicken IgY H&L (HRP)
    See all IgY secondary antibodies
  • Host species

    Goat
  • Target species

    Chicken
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Chicken IgY and with light chains common to other Chicken immunoglobulins. No antibody was detected against non immunoglobulin serum proteins.
  • Tested applications

    Suitable for: ICC, IHC-P, ELISA, WBmore details
  • Conjugation

    HRP

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    pH: 6.8
    Constituents: 0.2% BSA, 0.05% CMIT/MIT based preservative
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP).
  • Conjugation notes

    Molar enzyme/ antibody protein ratio is 4:1
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • General notes

    Part of the AbExcel range.
  • Research areas

    • Secondary antibodies
    • anti-Chicken
    • IgY
    • Enzyme
    • HRP

Associated products

  • Substrate reagent

    • TMB ELISA Substrate (Highest Sensitivity) (ab171522)
    • TMB ELISA Substrate (High Sensitivity) (ab171523)
    • TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    • TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
    • TMB ELISA Substrate (Slower Kinetic Rate) (ab171526)
    • TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab97135 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
ICC
Use at an assay dependent dilution.
IHC-P
1/200 - 1/5000.
ELISA (1)
1/10000 - 1/100000. (Primary).
WB (1)
1/2000 - 1/20000. Colorimetric: 1/5000 - 1/30000; Chemiluminescent: 1/10000 - 1/50000
Notes
ICC
Use at an assay dependent dilution.
IHC-P
1/200 - 1/5000.
ELISA
1/10000 - 1/100000. (Primary).
WB
1/2000 - 1/20000. Colorimetric: 1/5000 - 1/30000; Chemiluminescent: 1/10000 - 1/50000

Images

  • Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
    Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
    All lanes : Anti-beta Actin antibody - Loading Control (ab13822) at 1 µg/ml

    Lanes 1-6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1 - 2 : Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) (ab97135) at 1/2000 dilution
    Lanes 3 - 4 : Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) (ab97135) at 1/10000 dilution
    Lanes 5 - 6 : Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) (ab97135) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time : 90 seconds
  • Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
    Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)This image is courtesy of an anonymous Abreview.
    All lanes : Anti-Vimentin antibody (ab24525)

    All lanes : Mouse cardiac fibroblasts

    Lysates/proteins at 4 µg per lane.

    Secondary
    All lanes : Goat Anti-Chicken IgY H&L (HRP) (ab97135) at 1/2500 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 2 seconds

    See Abreview

  • Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)
    Western blot - Goat Anti-Chicken IgY H&L (HRP) (ab97135)

    Ab97135 used to detect a chicken anti-beta Galactosidase antibody (ab9361). Ab97135 used at a concentration of 1:2000 in 5% milk blocking buffer in 0.1% Tween, 1x PBS. Whole lysates from DH5a (E.coli). Specific band at approx. 110 KDa. Non-specific bands: 70 Kda. NB. Gels run on 10% Acrylamide gels.

    This image is courtesy of an anonymous Abreview.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (24)

Publishing research using ab97135? Please let us know so that we can cite the reference in this datasheet.

ab97135 has been referenced in 24 publications.

  • Uematsu K  et al. Osteoclastogenic Potential of Tissue-Engineered Periosteal Sheet: Effects of Culture Media on the Ability to Recruit Osteoclast Precursors. Int J Mol Sci 22:N/A (2021). PubMed: 33671612
  • Brady NJ  et al. Temporal evolution of cellular heterogeneity during the progression to advanced AR-negative prostate cancer. Nat Commun 12:3372 (2021). PubMed: 34099734
  • Chen P  et al. Structural basis for CSPG4 as a receptor for TcdB and a therapeutic target in Clostridioides difficile infection. Nat Commun 12:3748 (2021). PubMed: 34145250
  • Mason B  et al. Fbxl17 is rearranged in breast cancer and loss of its activity leads to increased global O-GlcNAcylation. Cell Mol Life Sci 77:2605-2620 (2020). PubMed: 31560077
  • Taglialegna A  et al. The biofilm-associated surface protein Esp of Enterococcus faecalis forms amyloid-like fibers. NPJ Biofilms Microbiomes 6:15 (2020). PubMed: 32221298
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-6 of 6 Abreviews or Q&A

ELISA with chicken plasma

Excellent
Abreviews
Abreviews
Application
ELISA
We followed the protocol published by Meunier et al. PLOS One 2017, after confirming with customer service that the concentration as published is the same for our lot number. The feed-back was fast and useful, and the ELISA results look good. We continue to use the product in our following ELISAs and are satisfied with the ease of use and the variability of the results.
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

Submitted Dec 20 2019

Rat astrocytes WB

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Antibody was diluted 1/2000 in TBST and incubated 1h at RT
Detection performed using ECL+
Primary antibody used was ab13989.
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

Submitted Oct 01 2015

Question

i am using this antibody for WB in yeast, would the GFP have any cross-reactivity in yeast to cause high background?

Read More

Abcam community

Verified customer

Asked on May 29 2013

Answer

Hello! The reply from the lab is below, please let me know if you have any further questions!



Two responses: The first is that the client should NOT use PVDF or any other nylon-based membranes. Chicken antibodies bind non-specifically to PVDF. Instead, they should use nitrocellulose membranes. Many folks aren't aware of this issue.



The second response is that there should be not cross-reactivity to anything in Saccharomyces or any other type of yeast. There are no yeast gene products that I am aware of that are similar to GFP. With that said, the yeast cell wall is tricky, and some things bind to it non-specifically (i.e., via ionic bonds with low affinity). Theoretically, these non-specific interactions can be abrogated with high salt. So, if the client is having trouble with higher-than-normal backgrounds (using nitrocellulose), then they should try making the total salt concentration 1.0 M in the primary antibody preparation.

Read More

Abcam Scientific Support

Answered on May 29 2013

Question

Das verwendete Kit wurde schon mehrmals getestet. Es funktioniert einwandfrei!
Gerne können wir den Antikörper-Test so machen wie Sie es vorschlagen!
Mit freundlichen Grüßen,

Read More

Abcam community

Verified customer

Asked on Nov 28 2012

Answer

Vielen Dank für Ihre Antwort.


Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie besprochen, habe ich eine kostenlose Ersatzlieferung des ab97135 für Sie in Auftrag gegeben. Sie hat die Referenznummer 1212315 und sollte am 30/11/12 bei Ihnen ankommen.

Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden wenn Sie weiterhin Probleme bei der Detektion haben und ich werde wie besprochen ein Austauschprodukt für den primären Antikörper veranlassen.

Read More

Abcam Scientific Support

Answered on Nov 28 2012

Question

leider haben wir im ganzen Haus keinen weiteren primären chicken Antikörper, das heißt, ich kann den sekundären Antikörper nicht einzeln testen.

Als Gewebe habe ich Maus Darm verwendet der auf jeden Fall LacZ positiv ist (wurde kolorimetrisch nachgewiesen).



Das von Ihnen empfohlene Protokoll habe ich schon ausprobiert, hat leider auch nicht funktioniert!



Vielen Dank,

Read More

Abcam community

Verified customer

Asked on Nov 27 2012

Answer

Vielen dank für Ihre Antwort.

In diesem Fall würde ich Ihnen gerne anbieten nach dem Ausschlussverfahren vorzugehen, wenn Sie damit einverstanden sind.

Ich würde Ihnen gerne ein kostenloses Ersatzprodukt für den sekundären Antikörper zusenden. Wenn Sie diesen für das Experiment verwenden und sie weiterhin kein positives Ergebnis erhalten würde ich im Anschluss gerne den primären Antikörper austauschen.

Es wäre hilfreich, wenn Sie in das nächste Experiment auch eine Kontrolle für das verwendete Kit mit einschließen könnten, um dieses auf die korrekte Funktionsweise hin zu überprüfen.

Bitte lassen Sie mich wissen was Sie von diesem Vorschlag halten. Sobald ich Ihre Rückmeldung erhalte werde ich die kostenlose Lieferung des Ersatzproduktes für den sekundären Antikörper gerne in Auftrag geben.

Read More

Abcam Scientific Support

Answered on Nov 27 2012

Question

anbei das ausgefüllte Protokoll für beide Antikörper. Vielen Dank schonmal im Voraus für Ihre Bemühungen!
Herzliche Grüße und schönes Wochenende,

Read More

Abcam community

Verified customer

Asked on Nov 23 2012

Answer

Vielen Dank für die Email und die Zusendung des ausgefüllten Fragebogens. Anja ist leider außer Haus. Wenn es Ihnen recht ist würde ich mich gerne in dieser Zeit um Ihren Fall kümmern.

Es tut mir Leid, dass sie Probleme mit unseren Produkten haben. Ich möchte Ihnen noch einmal versichern, dass der ab9361 garantiert ist für die ICH-Fr in Maus und damit unter unsere 6 Monatige Garantie fällt, wenn er nicht funktioniert wie auf dem Datenblatt beschrieben.

Leider gibt es nicht viele Veränderungen die ich zu Ihrem Protokoll vorschlagen kann. Um Ihnen zu helfen mehr Informationen zu erhalten und um zu Überprüfung, welcher der beiden Antikörper nicht funktioniert, moechte ich folgende Vorschläge machen.

1. Es wäre wichtig den zweiten Antikörper mit einem anderen primären Antikörper in einem Versuch zu verwenden, um dessen Funktionalität zu bestätigen.

2. Könnten Sie mir bitte genauerer Angaben über die verwendete positiv Kontrolle machen? Welches Gewebe wurde genau getestet?

3. Könnten Sie mir bitte genauere Angaben über das TSA Kit zukommen lassen. Die Herstellerangaben und eine Produktnummer wären hilfreich, so dass ich mir das Kit genauer anschauen kann.

4. Das von uns empfohlene Protokoll für die Färbung lautet wie folgt:

Day 1

1. (If using an HRP conjugate for detection, blocking of endogenous peroxidase can be performed here but

we recommend waiting until after the primary antibody incubation. See Day 2, step 2 and note v.).

2. Wash the slides 2 x 5 minutes in TBS plus 0.025% Triton X-100 with gentle agitation. (See note i).

3. Block in 10% normal serum with 1% BSA in TBS for 2 hours at room temperature (See note iii).

4. Drain slides for a few seconds (do not rinse) and wipe around the sections with tissue paper (See note iv).

5. Apply primary antibody diluted in TBS with 1% BSA (See note v).

6. Incubate overnight at 4°C (See note iv).

Day 2

1. Rinse 2 x 5min TBS 0.025% Triton with gentle agitation.

2. If using an HRP conjugate for detection, incubate the slides in 0.3% H2O2 in TBS for 15 min (See note v.).

3. For enzymatic detection (HRP or AP secondary conjugates):

Apply enzyme-conjugated secondary antibody to the slide diluted to the concentration recommended by

the manufacturer in TBS with 1% BSA, and incubate for 1 hour at room temperature.

For fluorescent detection:

4. Rinse 3 x 5min TBS.

If using fluorescent detection, end at this step and coverslip with mounting medium.

If visualizing the protein with a chromogen, continue with the following steps.

5. Develop with chromogen for 10 min at room temperature (See note vi.).

6. Rinse in running tap water for 5 min.

7. Counterstain (if required) (See note vii.).

8. Dehydrate, clear and mount (See notes ix. and x.).

Bitte zögern Sie nicht mit den zusätzlichen Informationen auf mich zurück zu kommen. Der vermutlich wichtigste erste Schritt ist zu überprüfen welcher der beiden Antikörper nicht funktioniert.

Ich hoffe ich konnte Ihnen etwas weiter helfen und freue mich auf Ihre Antwort.

Read More

Abcam Scientific Support

Answered on Nov 23 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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