YILIMART - KIR Typing Kit (24 tests)

The KIR Typing Kit allows the detection of human killer cell immunoglobulin-like receptors (KIRs) on genomic DNA level or the analysis of KIR expression on mRNA level.

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The presence or absence of KIR genes is analyzed by PCR using carefully designed sequence-specific primers (SSPs), enabling the detection of all known 15 human KIR genes plus two pseudo genes. Allel coverage: IPD KIR Sequence database release 2.1, March 2009 (http://www.ebi.ac.uk/ipd/)

Each well of the 96-well PCR plate contains a lyophilized enzyme mix, including Taq DNA Polymerase, as well as positive control primers. Resuspension Buffer is simply mixed with the template and dispensed into the wells of the PCR plate. PCR products are ready for immediate analysis by electrophoresis due to the integrated gel loading buffer.

Background information

KIRs can be found on natural killer (NK) cells which play an important role in the innate immune response. NK cells have been implicated in the suppression of graft-versus-host disease, promotion of bone marrow engraftment, and mediation of a graft-versus-leukemia effect. KIRs exist in various isoforms. The inhibitory KIRs interact with specific HLA class I molecules, predominantly HLA-C, on target cells.

Not all KIR genes are present in every human individual. Further heterogeneity exists at the transcriptional level. Different subsets of NK cells express different KIRs, even within one individual.

Downstream applications

The KIR Typing Kit allows the investigation of the KIR genotype on the DNA level and its expression on mRNA level. The KIR Typing Kit has been used, for example, for KIR profiling after haploidentical stem cell transplantation and for research into the role of KIR genotypes in susceptibility to infectious disease.

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Figure 1
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Figure 1

KIR Typing Kit
with ready-to-use solutions, lyophilized enzyme mix in a 96-well PCR plate, and protocol.

KIR Typing Kit
with ready-to-use solutions, lyophilized enzyme mix in a 96-well PCR plate, and protocol.
Figure 2
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Figure 2

KIR genotyping

Agarose gel separation of PCR products obtained from genomic DNA isolated from whole blood. Lanes 1–5, 7, 9–12, 14–16, and 18–19 show positive KIR (+). Lanes 6, 8, 13, and 17 are negative for KIR (–).

In addition all lanes show positive control bands (400 bp). Lane 20 shows the genomic DNA control band at 260 bp, lane 21 the positive control, and lane 22 the negative control. M: DNA marker

KIR genotyping

Agarose gel separation of PCR products obtained from genomic DNA isolated from whole blood. Lanes 1–5, 7, 9–12, 14–16, and 18–19 show positive KIR (+). Lanes 6, 8, 13, and 17 are negative for KIR (–).

In addition all lanes show positive control bands (400 bp). Lane 20 shows the genomic DNA control band at 260 bp, lane 21 the positive control, and lane 22 the negative control. M: DNA marker
Figure 3
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Figure 3

mRNA expression profiling of KIR genes

Agarose gel separation of PCR products obtained from mRNA-derived cDNA isolated from 5 mL whole blood. Lanes 1–4, 9–12 and 14–15 show positive KIR bands (+), whereas lanes 5–8, 13, and 16–19 are negative for KIR (–).

In addition, all lanes show positive control bands (400 bp). Lane 20 shows no genomic DNA control band at 260 bp; thus, there is no genomic DNA contamination. M: DNA marker

mRNA expression profiling of KIR genes

Agarose gel separation of PCR products obtained from mRNA-derived cDNA isolated from 5 mL whole blood. Lanes 1–4, 9–12 and 14–15 show positive KIR bands (+), whereas lanes 5–8, 13, and 16–19 are negative for KIR (–).

In addition, all lanes show positive control bands (400 bp). Lane 20 shows no genomic DNA control band at 260 bp; thus, there is no genomic DNA contamination. M: DNA marker
Figure 4
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Figure 4

Gene name and allele specificity

The KIR Typing Kit identifies the presence or absence of the following KIR genes and variants.

(Allel coverage: IPD KIR Sequence database release 2.1, march 2009 (http://www.ebi.ac.uk/ipd/)

Gene name and allele specificity

The KIR Typing Kit identifies the presence or absence of the following KIR genes and variants.

(Allel coverage: IPD KIR Sequence database release 2.1, march 2009 (http://www.ebi.ac.uk/ipd/)

Selected references
1. Basu, D. et al. (2009) Stimulatory and inhibitory killer Ig-like receptor molecules are expressed and functional on lupus T cells. J. Immunol. 183: 3481-3487
2. Pérez-Martínez, A. et al. (2009) KIR-HLA receptor-ligand mismatch associated with a graft-versus-tumor effect in haploidentical stem cell transplantation for pediatric metastatic solid tumors. Pediatr. Blood Cancer 53: 120-124
3. André, M. C. et al. (2010)
  
  Long-term human CD34
  ⁺
  stem cell-engrafted nonobese diabetic/SCID/IL-2R gamma(null) mice show impaired CD8
  ⁺
  T cell maintenance and a functional arrest of immature NK cells.
  
  J. Immunol. 185: 2710-2720
4. Siegler, U. et al. (2010) Good manufacturing practice-compliant cell sorting and large-scale expansion of single KIR-positive alloreactive human natural killer cells for multiple infusions to leukemia patients. Cytotherapy 12(6): 750-763
5. Wauquier, N. et al. (2010) Association of KIR2DS1 and KIR2DS3 with fatal outcome in Ebola virus infection. Immunogenetics 62: 767-771
Brochures and posters

KIR typing
Customer reports and application notes

Cross-reactivity of KIR antibodies

Flow cytometry analysis of whole-blood NK cells expressing single killer cell immunoglobulin–like receptors
Special protocols

cDNA synthesis from whole blood using the μMACS cDNA Kit and subsequent KIR typing
Further downloads

KIR Typing Kit – Evaluation Form

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KIR Typing Kit (24 tests)

For research use only

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Background information

Downstream applications

Figure 1

Figure 1

Figure 2

Figure 2

Figure 3

Figure 3

Figure 4

Figure 4

Selected references

Brochures and posters

Customer reports and application notes

Special protocols

Further downloads

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