Overview

  • Product name
    Anti-MHC Class I H2 Kb antibody
  • Description
    Rabbit polyclonal to MHC Class I H2 Kb
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Recombinant fragment
  • Immunogen

    Synthetic peptide corresponding an internal sequence of Mouse MHC Class I H2 Kb.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 50% Glycerol, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purity >90%
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab93364 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 41 kDa. This antibody has been tested in Western blot against the recombinant peptide used as an immunogen. We have no data on detection of endogenous protein.
ELISA 1/2000 - 1/5000.

Target

  • Relevance
    H2-Kb belongs to the family of the mouse Major Histocompatibility Complex class I receptors. These display processed antigens from the endocytosed bacteria. MHC class I proteins can form complexes with processed antigen peptides only if the antigen is synthesized in the same cell. As a consequence, T lymphocytes can only bind to class-I-positive cells that are synthesizing the antigen (e.g. virus-infected cells).
  • Cellular localization
    Cell Membrane
  • Database links
  • Alternative names
    • H2 class I histocompatibility antigen antibody
    • H2K antibody
    • H2K(B) antibody
    • H2K1 antibody
    • KB alpha chain antibody
    see all

Images

  • All lanes : Anti-MHC Class I H2 Kb antibody (ab93364) at 1 µg/ml

    Lane 1 : Whole cell lysate prepared from murine insulinoma cells MIN6 untreated.
    Lane 2 : Whole cell lysate prepared from murine insulinoma cells MIN6 treated with 60U/ml interferon-gamma for 72 hours.

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP conjugated goat anti-rabbit (H+L) polyclonal at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 41 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 seconds

    See Abreview

References

This product has been referenced in:
  • Maschalidi S  et al. UNC93B1 interacts with the calcium sensor STIM1 for efficient antigen cross-presentation in dendritic cells. Nat Commun 8:1640 (2017). Read more (PubMed: 29158474) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Colon Cancer Cell Line- CT26)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
30 µg
Specification
Mouse Colon Cancer Cell Line- CT26
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Aug 19 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - 0.1M PBS with 3% TritonX100
Fixative
Paraformaldehyde

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Nov 05 2014

Application
Western blot
Loading amount
1.2e+007 cells
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Mouse Tissue lysate - whole (spleenocytes)
Specification
spleenocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 17 2014

Answer

we have issued a free of charge replacement.

Read More

Question

1) Abcam product code ab ab93364

2) Abcam order reference number or product batch number

Rabbit polyclonal to MHC Class I H2-Kb

GR101536

3) Description of the problem / absence of staining

4) Sample preparation:

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

Lysis buffer NP40 1%

Protease inhibitors: cocktail from Roche

Phosphatase inhibitors no

Reducing agent SDS

Boiling for ≥5 min? yes

Protein loaded ug/lane or cells/lane 50

Positive control yes

Negative control no



5) Percentage of gel 12%

Type of membrane PVDF

Protein transfer verified yes

Blocking agent and concentration tried both BSA and skim milk 5%

Blocking time 1 hour

Blocking temperature room temperature



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 1 ug/ml

Diluent buffer blocking buffer

Incubation time overnight tried room temperature one hour

Incubation temperature: 4C



7) Secondary antibody:

Species: goat anti rabbit

Reacts against:

Concentration or dilution 1/25 000

Diluent buffer blocking buffer

Incubation time room temoperature

Incubation temperature: one hour

Fluorochrome or enzyme conjugate: HRP



8) Washing after primary and secondary antibodies:

Buffer PBS-Tween

Number of washes 4



9) Detection method ECL



10) How many times have you run this staining? 4

Do you obtain the same results every time? yes

What steps have you altered to try and optimize the use of this antibody? Lysis buffer //Blocking buffer // time of incubation



ATTACHED is a picture of the actin from one of the blot done with abcam antibody and on the left the exon8 antibody specific for MHC-class 1 H2Kb from a collaborator

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Answer

Thank you for contacting us.

Your credit note ID is 23807.

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

Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Could you provide some further details of the protocol used and complete the following form (attached as a word document).

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
Application
Western blot
Sample
Mouse Cell lysate - whole cell (murine insulinoma cells MIN6)
Loading amount
25 µg
Specification
murine insulinoma cells MIN6
Treatment
60U/ml interferon-gamma for 72hours
Gel Running Conditions
Reduced Denaturing (10% Resolving gel, 5% stacking gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Apr 19 2012

Question
Answer

DISCOUNT CODE: xx Ablaufdatum: 20. Juli 2012

Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper ab93364 im Western blot gegen das endogene Mausprotein zu testen, annehmen möchten. Wie am Telefon besprochen, können Sie den Gutscheincode zum Kauf eines weiteren Antikörpers einsetzen, sobald Sie uns ein Abreview für diesen Antikörper eingereicht haben. Der Discountcode hat die Nummer xx, deckt den Wert von ab93364 ab und kann zum Kauf weiterer Produkte eingesetzt werden.

Der Code wird gültig, sobald Sie uns ein Abreview (www.abcam.com/Abreviews) für diesen Antikörper über den Test gegen das endogene Mausprotein eingereicht haben. Bitte geben Sie diesen Code auch im Abschnitt “Additional Notes” des Abreviews mit an, so dass wir wissen, dass sich dieses Abreview auf die Gutscheinaktion bezieht. Der Code wird dadurch aktiviert. Bitte kontaktieren Sie bei der nächsten Bestellung unsere Kundendienst mit dem Code und der ursprünglichen Bestellnummer.

Bitte zögern Sie nicht, sich wieder an uns zu wenden, falls Sie weitere Fragen haben. Wir freuen uns auf Ihr Abreview, egal ob Ihre Ergebnisse positiv oder negativ sind und wünschen Ihnen viel Glück für Ihre Forschung.

Die Bedingungen dieses Angebotes befinden sich unter dem folgenden Link: www.abcam.com/collaborationdiscount


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Answer

Thank you for contacting us. We have only tested this antibody with recombinant fragment which we used as immunogen. We have not tested it using endogenous protein or lysates. This is the reason why the mouse species is predicted not tested. Abpromise does not apply to the species that are predicted. However this product is eligible for Abreview discount with mouse species. Let me know if the customer is interested in Abreview discount offer. Unfortunately we haven’t done any research on which MHC allele the C57B6 mouse displays so I would recommend doing search in literature like below  http://www.pnas.org/content/early/2011/02/02/1005342108.full.pdf I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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Answer

Thank you for contacting us. I can confirm that using this MHC Class I H2 Kb antibody ab93364 a lower band can be expected at around 40 kDa which is due to the non-glycosylated form of the protein. The upper band is the glycosylated mature protein. The predicted molecular weight and glycosylation information can be found on the SwissProt link for this protein: http://www.uniprot.org/uniprot/P01901 Unfortunately, we do not have any comments from other users since May. However, we would be very grateful if you could submit an Abreview about your experience with this product on endogenous protein samples via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to researchers. To find out more about our Abreview system, please see the following webpage: https://www.abcam.com/abreviews I hope this information helps. Please do not hesitate to contact us if you need anything further.

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1-10 of 11 Abreviews or Q&A

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