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Antibodies

CIITA Antibody

Product Sizes
100 ul
100-401-249-100UL
About this Product
SKU:
100-401-249
Additional Names:
CIITA|rabbit anti-CIITA Antibody, MHC class II transactivator, CIITA, MHC2TA
Application:
ChIP, ELISA, Immunoprecipitation, Western Blot
Buffer:
None
CE/IVD:
RUO
Clonality:
Polyclonal
Concentration:
90 mg/ml
Extra Details:
Anti-CIITA antibody detects CIITA. The transactivator CIITA regulates basal and interferon-induced expression of Major Histocompatibility Complex class II genes. CIITA restores expression of all MHC class II gene expression in mutant cells and corrects regulatory defects of MHC class II genes. Antibodies to this transactivator are useful in the study of diseases of pathological MHC class II expression. Antigen can be obtained from Raji cell lysates. Typically levels of CIITA expression are too low to detect endogenous levels of protein expression. Transiently transfected cells are usually employed to study this transcription factor.
Format:
Antiserum
Host:
Rabbit
Immunogen:
The immunogen used for this study was a bacterially produced recombinant FLAG-CIITA corresponding to amino acids 1 through 333 of the human protein.
Physical State:
Liquid
Reactivities:
Human
Shipping Conditions:
Dry Ice
Specificity:
Anti-CIITA antibody was prepared from monospecific antiserum by delipidation and defibrination.
Storage Conditions:
-20[o]C aliquoted. Aliquot. Avoid freeze/thaw cycles.
Supplier:
Rockland Inc
Type:
Antibody: Polyclonal Antibody
Western blot of Anti-CIITA (1-333) antibody, generated by immunization with bacterially produced FLAG-CIITA aa 1-333, was tested by western blot against lysates of Cos-7 cells after transient transfection, separately, with pcDNA3-FLAG-CIITA-336-884 and pcDNA3-HA-CIITA. For transfection, Fugene 6 (Roche) was used according to the manufacturer's instructions for a 6-well plate format. Cells were lysed 24 h post-transfection in 200 uL of 1x SDS-sample buffer, heated at 96°C for 5', and vortexed for 30 sec. Samples (10 uL each) were separated on a 12% SDS-PAGE gel and transferred to PVDF (Millipore) followed by blocking for 45' using TTBS supplemented with 5% non-fat dry milk. All incubations were performed at room temperature. In panel A, both samples on PVDF were incubated with 10 ug/mL mouse anti-FLAG antibody (Sigma) for 45'. After 5X washes with TTBS, reaction with ALP rabbit anti-mouse IgG at 200 ng/mL proceeded for 45' following again by washing as before. The blot was developed using BCIP/NBT. This blot demonstrates that FLAG-CIITA-336-884 was successfully over-expressed in the Cos-7 cells. In panel B, both samples on PVDF were incubated with a 1:500 dilution of Rockland's anti-CIITA (1-333) for 45'. After 5X washes with TTBS, reaction with HRP goat anti-rabbit IgG at 10 ng/mL proceeded for 45' following again by washing as before. The membrane was covered with Pico West Substrate solution (Pierce) for 5' and was then placed between the two layers of a standard sheet protector. Kodak O-MAT film was exposed to the blot for 30 sec and was immediately developed. The lane containing the lysate of pcDNA3-HA-CIITA transfected cells contains a single band at ~130 kDa molecular weight, whereas the lane containing lysate from pcDNA3-FLAG-CIITA-336-884 transfected cells shows no reactivity. This blot demonstrates that anti-CIITA (1-333) is specific for amino acids 1-333 of CIITA and that the antibody is not cross reactive with the FLAG portion of the immunogen.