Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Rat (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
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Product Specifications

Immunogen

A synthetic peptide made to an internal portion of the human Zic1 protein (between residues 20-75) [UniProt Q15915]

Reactivity Notes

Predicted to react with rat based on 100% sequence homology.

Localization

Nuclear.

Marker

Ectoderm Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for ZIC1 Antibody - BSA Free

Western Blot: ZIC1 AntibodyBSA Free [NB600-488]

Western Blot: ZIC1 AntibodyBSA Free [NB600-488]

Western Blot: ZIC1 Antibody [NB600-488] - WB analysis of Zic1 in overexpression lysate.
Immunohistochemistry-Paraffin: ZIC1 Antibody - BSA Free [NB600-488]

Immunohistochemistry-Paraffin: ZIC1 Antibody - BSA Free [NB600-488]

ZIC1-Antibody-Immunohistochemistry-Paraffin-NB600-488-img0005.jpg
Western Blot: ZIC1 AntibodyBSA Free [NB600-488]

Western Blot: ZIC1 AntibodyBSA Free [NB600-488]

Western Blot: ZIC1 Antibody [NB600-488] - Western blot analysis of Zic1 in mouse cerebellum extract. Protein extracts were prepared from mouse cerebellum between postnatal day 5 (P5) and P18, as indicated above the lanes.
Immunohistochemistry: ZIC1 Antibody - BSA Free [NB600-488]

Immunohistochemistry: ZIC1 Antibody - BSA Free [NB600-488]

Immunohistochemistry: ZIC1 Antibody [NB600-488] - IHC analysis of Zic1 in mouse kidney using DAB with hematoxylin counterstain.
Immunohistochemistry-Paraffin: ZIC1 Antibody - BSA Free [NB600-488]

Immunohistochemistry-Paraffin: ZIC1 Antibody - BSA Free [NB600-488]

ZIC1-Antibody-Immunohistochemistry-Paraffin-NB600-488-img0006.jpg
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Altered brain pericyte proliferation in Foxc1 mutants.(A–C) PDGFr beta (green) & IB4 (red) immunofluorescence highlight pericyte cell bodies in the cortices of E14.5 WT, Foxc1h/h & Foxc1l/l. (D–F) Triple immunofluorescence for Zic1 (pericyte nuclei; green), BrdU (red) & IB4 (blue) highlight proliferating (Zic1+/BrdU+; arrows) pericytes in the E14.5 cerebral vasculature of WT, Foxc1h/h & Foxc1l/l embryos. (G) Graph depicting quantification of Zic1+ pericyte nuclei/vessel length in all three genotypes at E14.5. (H) Graph depicting quantification of pericyte proliferation (labeling index) at E14.5. Scale bars: 100 µm. Means represent analysis of three independent litters for each mutant genotype (n = 3). Asterisks indicate a statistically significance difference from WT (P<0.05). Error bars depict ±SEM. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Altered brain pericyte proliferation in Foxc1 mutants.(A–C) PDGFr beta (green) & IB4 (red) immunofluorescence highlight pericyte cell bodies in the cortices of E14.5 WT, Foxc1h/h & Foxc1l/l. (D–F) Triple immunofluorescence for Zic1 (pericyte nuclei; green), BrdU (red) & IB4 (blue) highlight proliferating (Zic1+/BrdU+; arrows) pericytes in the E14.5 cerebral vasculature of WT, Foxc1h/h & Foxc1l/l embryos. (G) Graph depicting quantification of Zic1+ pericyte nuclei/vessel length in all three genotypes at E14.5. (H) Graph depicting quantification of pericyte proliferation (labeling index) at E14.5. Scale bars: 100 µm. Means represent analysis of three independent litters for each mutant genotype (n = 3). Asterisks indicate a statistically significance difference from WT (P<0.05). Error bars depict ±SEM. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Pericyte conditional Foxc1 mutants have increased pericyte proliferation & density in the cerebral vasculature.(A,C) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. (B,D) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E14.5. Arrowheads indicate PDGFr beta + pericytes. (E,F) Zic1 (green), BrdU (red), & IB4 (blue) triple immunofluorescence of E14.5 PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ cerebral cortex. Arrows indicate Zic1+/BrdU+ pericytes. (G) Graph depicts quantification of Zic1+ pericyte proliferation (labeling index) in the cerebral vasculature at E14.5. (H,J) Zic1 (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. (I,K) PDGFr beta (green) & IB4 (red) co-immunofluorescence in the cortices of PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ brains at E18.5. Arrowheads indicate PDGFr beta + pericytes. (L) Graph depicts pericyte density (Zic1+ pericyte nuclei/vessel length) in the cerebral vasculature at E14.5 & E18.5 in PDGFr beta -cre; Foxc1fl-fl mutant & PDGFr beta -cre; Foxc1fl-+ animals. Asterisks indicate a statistically significance difference from PDGFr beta -cre; Foxc1fl-+ samples (P<0.05). Analysis of pericyte proliferation & density was performed on control & mutant brains from three separate litters at each time point (n = 3). Error bars depict ±SEM. Scale bars in A,C,E,F,H,J: 100 µm. Scale bars in B,D,I,K: 50 µm. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ZIC1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] -

Immunocytochemistry/ Immunofluorescence: ZIC1 Antibody - BSA Free [NB600-488] - Altered brain pericyte proliferation in Foxc1 mutants.(A–C) PDGFr beta (green) & IB4 (red) immunofluorescence highlight pericyte cell bodies in the cortices of E14.5 WT, Foxc1h/h & Foxc1l/l. (D–F) Triple immunofluorescence for Zic1 (pericyte nuclei; green), BrdU (red) & IB4 (blue) highlight proliferating (Zic1+/BrdU+; arrows) pericytes in the E14.5 cerebral vasculature of WT, Foxc1h/h & Foxc1l/l embryos. (G) Graph depicting quantification of Zic1+ pericyte nuclei/vessel length in all three genotypes at E14.5. (H) Graph depicting quantification of pericyte proliferation (labeling index) at E14.5. Scale bars: 100 µm. Means represent analysis of three independent litters for each mutant genotype (n = 3). Asterisks indicate a statistically significance difference from WT (P<0.05). Error bars depict ±SEM. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/2/7/647/845/Foxc1-is-requir…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for ZIC1 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:400

Immunohistochemistry-Frozen

reported in scientific literature (PMID 23862012)

Immunohistochemistry-Paraffin

1:400

Western Blot

1:1000
Application Notes
Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.01% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: ZIC1

Zinc finger protein ZIC 1 (ZIC1) is a member of the ZIC family of C2H2-type zinc finger proteins. These proteins are particularly important during embryonic development. ZIC1 is a transcriptional activator via interaction with GLI1, GLI2, and GLI3. ZIC1 facilitates development and maturation of the cerebellum and spinal cord during organogenesis of the CNS. Within the pontine gray nucleus (PGN) ZIC1 plays a role in spatial distribution of mossy fiber neurons. The interaction between Zic and Gli family proteins are significant for molecular antagonistic and synergistic factors in development. Zic has been shown involvement in modulation of the hedgehog signaling pathway. ZIC1 may be found in the nucleus or in the cytoplasm in the presence of MDFIC overexpression. This protein is mainly found within the CNS and has high levels of expression in the cerebellum. In cancerous tissue, the ZIC1 protein is often found in medulloblastomas. ZIC1 has been used as aneuroepithelial marker.

Long Name

Zic Family Member 1

Alternate Names

ZNF201

Entrez Gene IDs

7545 (Human); 22771 (Mouse); 64618 (Rat)

Gene Symbol

ZIC1

UniProt

Additional ZIC1 Products

Product Documents for ZIC1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for ZIC1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for ZIC1 Antibody - BSA Free

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Protocols

View specific protocols for ZIC1 Antibody - BSA Free (NB600-488):

ZIC1 Antibody:
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

ZIC1 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for ZIC1 Antibody - BSA Free

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  • Q: Has this antibody been recently qualified for use in chromatin immunoprecitiation?

    A: NB600-488 has not been validated and can therefore not be guaranteed for ChIP. If you would be interested in testing this novel application, please take a look at our Innovators Reward Program.

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