Oct4 Recombinant Antibody

Lane 1: F9; Lane 2: NCCIT; Lane 3: hES lysates probed with Oct 4 (2D5) Monoclonal Antibody (bsm-52002R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.

N2A cells were stained with Oct 4 (2D5) Monoclonal Antibody (bsm-52002R) at [1:200] incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$Western blot analysis of Oct4 on F9 cell lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/1,000 dilution.\r\n\r\nLysates\/proteins at 10 \u788cg\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 2 minutes;\r\n\r\n12% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) at 1\/1,000 dilution was used in 5% NFDM\/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.\r\n$

Western blot analysis of Oct4 on F9 cell lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/1,000 dilution.\r\n\r\nLysates\/proteins at 10 \u788cg\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 2 minutes;\r\n\r\n12% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) at 1\/1,000 dilution was used in 5% NFDM\/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.\r\n

$Western blot analysis of Oct4 on different lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/500 dilution.\r\n\r\nLane 1: HES cell lysate\r\nLane 2: NCCIT cell lysate\r\n\r\nLysates\/proteins at 10 \u00b5g\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 1 minute;\r\n\r\n10% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) at 1\/500 dilution was used in 5% NFDM\/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.\r\n$

Western blot analysis of Oct4 on different lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/500 dilution.\r\n\r\nLane 1: HES cell lysate\r\nLane 2: NCCIT cell lysate\r\n\r\nLysates\/proteins at 10 \u00b5g\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 1 minute;\r\n\r\n10% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) at 1\/500 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.\r\n

$ICC staining of Oct4 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-52002R, 1\/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor\u00ae488 Goat anti-Rabbit IgG was used as the secondary antibody at 1\/1,000 dilution. The nuclear counter stain is DAPI (blue).$

ICC staining of Oct4 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-52002R, 1\/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor\u00ae488 Goat anti-Rabbit IgG was used as the secondary antibody at 1\/1,000 dilution. The nuclear counter stain is DAPI (blue).

$Immunohistochemical analysis of paraffin-embedded human seminoma tissue tissue with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/4,000 dilution.\r\n\r\nThe section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52002R) at 1\/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.$

Immunohistochemical analysis of paraffin-embedded human seminoma tissue tissue with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/4,000 dilution.\r\n\r\nThe 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 (bsm-52002R) at 1\/4,000 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.

Applications

WB
IHC-P
IHC-F
IF
ICC/IF

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52002r
Product Name	Oct4 Recombinant Antibody
Applications	WB, IHC-P, IHC-F, IF, ICC/IF
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human Oct 4 protein, around 1-100aa.
Clonality	Recombinant
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition	Shipped at 4°. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5460
Swiss Prot	Q01860
Synonyms	MGC22487; Oct 3; Oct 4; Oct-3; Oct-4; OCT3; Oct4; Octamer binding protein 3; Octamer-binding protein 3; Octamer binding protein 4; Octamer binding transcription factor 3; Octamer-binding transcription factor 3; Octamer-4; POU domain transcription factor OCT4; OTF3; OTF4; OTF 3; OTF 4; OTF-3; POU class 5 homeobox 1; POU domain, class 5, transcription factor 1; POU domain class 5 transcription factor 1; PO5F1_HUMAN; POU type homeodomain containing DNA binding protein ; POU5F1.
Background	Expression of the POU-domain transcription factor Octamer-4 (Oct-4) is widely regarded as a hallmark of pluripotent stem cells. The relationship of Oct-4 to pluripotent stem cells is indicated by its tightly restricted expression to undifferentiated pluripotent stem cells. Upon differentiation to somatic lineages, the expression of Oct-4 disappears rapidly. Unlike the majority of pluripotent stem cell markers, the biological role of Oct-4 has been well characterized. Studies performed in mice point to the critical role of Oct-4 in the establishment and/or maintenance of pluripotent stem cells in an uncommitted state.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC-F	1:100-500
IF
ICC/IF	1:50-100