HAK21032

iPS Cell Reprogramming Antibody Kit

cat.: HAK21032

Product Type:	Antibody Sampler Kit
Species reactivity:	Human
Applications:	WB

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Uniprot #:	SwissProt: Q01860 Human \| P20263 Mouse \| P48431 Human \| P48432 Mouse \| Q9H9Z2 Human \| O43474 Human \| Q60793 Mouse \| P01106 Human \| P01108 Mouse \| P09416 Rat Entrez Gene: 499593 Rat Unigene: 7719 Rat

Images

Fig1: All lanes: Western blot analysis of Oct4 with anti-Oct4 antibody (ET1612-20) at 1/500 dilution.

Lane 1: Wild-type NCCIT whole cell lysate.
Lane 2: Oct4 knockout NCCIT whole cell lysate.

ET1612-20 was shown to specifically react with Oct4 in Wild-type NCCIT cells. No band was observed when Oct4 knockout sample was tested. Wild-type and Oct4 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-Oct4 antibody (ET1612-20, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of NCCIT cells labeling Oct4 with Rabbit anti-Oct4 antibody (ET1612-20) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Oct4 antibody (ET1612-20) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with Oct4 (ET1612-20) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
	Fig4: Immunohistochemical analysis of paraffin-embedded human glioma tissue with Rabbit anti-SOX2 antibody (HA721155) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721155) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with SOX2 (HA721155) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Fig6: Western blot analysis of KLF4 on different lysates with Rabbit anti-KLF4 antibody (ET1702-71) at 1/2,000 dilution.

Lane 1: 293 cell lysate
Lane 2: NCCIT cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute 30 seconds;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-71) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig7: Western blot analysis of c-Myc on different lysates with Rabbit anti-c-Myc antibody (HA721182) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: Daudi cell lysate
Lane 3: K562 cell lysate
Lane 4: HL-60 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 55 kDa

Exposure time: 2 minutes;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721182) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.