Rabbit anti cdk4

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Rabbit anti-CDK4 is a primary antibody produced in rabbits that specifically binds to the Cyclin-Dependent Kinase 4 (CDK4) protein. CDK4 is a serine/threonine protein kinase that plays a crucial role in cell cycle regulation, particularly in the G1/S phase transition.

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Lab products found in correlation

Mouse anti cyclin d1, by Santa Cruz Biotechnology (2 mentions) Mouse anti gfap, by Merck Group (1 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Molecular imaging software, by Carestream (1 mentions) Rabbit anti iba1, by Fujifilm (1 mentions) Rabbit anti cyclin d1, by Cell Signaling Technology (1 mentions) Na934v, by Merck Group (1 mentions) β tubulin, by Abcam (1 mentions) Rabbit anti α tubulin, by Cell Signaling Technology (1 mentions) Complete ultra tablet, by Roche (1 mentions) Ripa buffer, by Merck Group (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Hrp conjugated against rat or rabbit, by Jackson ImmunoResearch (1 mentions) Bca assay, by Merck Group (1 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Rhodamine conjugated phalloidin, by Merck Group (1 mentions) Mouse anti stat3, by Abcam (1 mentions) Mouse anti ccnd1, by Santa Cruz Biotechnology (1 mentions) Mouse anti bcl 2, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Anti-CDK4 (57 citations) Rabbit anti-TM (2 citations) Rabbit anti-CDK4 (SC-260, (2 citations)

8 protocols using rabbit anti cdk4

Spinal Cord Immunofluorescence Staining

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Lumbar enlargement 4-5 was sectioned for immunofluorescence staining followed procedures described previously [61 (link)]. The following primary antibodies were used: mouse anti-cyclin D1 (1:500; Thermal); rabbit anti-CDK4 (1:500; Santa Cruz Biotechnology); rat anti-ED1 (1:500; AbD Serotec); rabbit anti-Ionized calcium binding adaptor molecule 1 (Iba-1, 1:1000, Wako Chemicals); rabbit anti-glial fibrillary acid protein (GFAP, 1:500, Millipore); and mouse anti-GFAP (1:500, Sigma). Images were acquired using a Leica TCS SP5 II Tunable Spectral Confocal microscope system (Leica Microsystems Inc) and processed using Adobe Photoshop 7.0 software (Adobe Systems). All immunohistological staining experiments were performed with appropriate positive control tissue as well as primary/secondary-only negative controls. For quantitative image analysis, digital images at 63× magnification were captured from both sides of spinal dorsal horns. The number of cyclin D1, CDK4, ED1/Iba-1, and GFAP labeled cells was quantified for each animal by a total # of both the left and right sides of dorsal horns (four images from 4 sections per mouse). All images were captured from n=5–6 mice per group.

Wu J., Zhao Z., Zhu X., Renn C.L., Dorsey S.G, & Faden A.I. (2016). Cell cycle inhibition limits development and maintenance of neuropathic pain following spinal cord injury. Pain, 157(2), 488-503.

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Spinal Cord Injury Western Blot Analysis

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Mouse spinal cord tissue (three-millimeter segments) centered at the injury site was obtained at indicated time for Western blot analysis performed as described previously [61 (link)]. Primary antibodies included: mouse anti-cyclin D1 (1:500; Thermal); rabbit anti-CDK4 (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, UAS); rabbit anti-Iba-1 (1:1000; Wako Chemicals), and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:2000; Millipore). Immune complexes were detected with the appropriate HRP conjugated secondary antibodies (KPL, Inc., Gaithersburg, MD) and visualized using SuperSignal West Dura Extended Duration Substrate (Thermo Scientific, Rockford, IL). Chemiluminescence was captured on a Kodak Image Station 4000R station (Carestream Health Inc., Rochester, NY) and protein bands were quantified by densitometric analysis using Carestream Molecular Imaging Software (Carestream Health Inc., Rochester, NY). The data presented reflects the intensity of target protein band compared to control and normalized based on the intensity of the endogenous control for each sample (expressed in fold of sham).

Wu J., Zhao Z., Zhu X., Renn C.L., Dorsey S.G, & Faden A.I. (2016). Cell cycle inhibition limits development and maintenance of neuropathic pain following spinal cord injury. Pain, 157(2), 488-503.

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Western Blot Analysis of Orai3, Cyclin D1, and Cdk4

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Total protein was extracted with NP40 lysis buffer along with protease inhibitor cocktail. Total protein concentration was quantified by BCA assay. Protein extracts were separated by 8–12% SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 5% skimmed milk in Tris-buffered saline with Tween 20 (TBST) for 2 h at room temperature, and then incubated overnight at 4 °C in TBST with primary antibody, including rabbit anti-Orai3 (1:500, Abcam), rabbit anti-cyclin D1 (1:1000, Cell Signaling) and rabbit anti-Cdk4 (1:1000, Santa Cruz). Following incubation with horseradish peroxidase-conjugated donkey anti-rabbit secondary antibody (1:5000, NA934V, Sigma, St. Louis, MO, USA) for 2 h, the membranes were detected using the enhanced chemiluminescence (ECL) kit. β-tubulin (1:5000, Abcam) was used as a loading control. Densitometric analysis was done using ImageJ software and data are graphically represented as mean ± SEM.

Arora S., Tanwar J., Sharma N., Saurav S, & Motiani R.K. (2021). Orai3 Regulates Pancreatic Cancer Metastasis by Encoding a Functional Store Operated Calcium Entry Channel. Cancers, 13(23), 5937.

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Western Blot Analysis of Angiogenic Factors

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Lysis was done with RIPA buffer (Sigma-Aldrich, St. Louis, MO, USA) containing protease and phosphatase inhibitors (Complete ULTRA Tablets, Roche, Nutley, NJ, USA). After centrifugation, the supernatant was collected and quantified by BCA assay (Sigma-Aldrich). The proteins were then separated by SDS-PAGE and transferred to nitrocellulose membranes. After blocking with 5% non-fat milk, the membranes were probed with rabbit anti-VEGF₁₆₅ (R&D System, Los Angeles, CA, USA), rabbit anti- VEGF₁₆₅b (R&D System), rabbit anti-SRPK1 (Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 (Cell Signaling Technology), rabbit anti-cleaved caspase 3 (casps3; Cell Signaling Technology), rabbit anti-CCND1 (Santa Cruz Biotechnology, Dallas, Texas, USA), rabbit anti-CDK4 (Santa Cruz Biotechnology), rabbit anti-nitric oxide synthase (anti-NOS3; Cell Signaling Technology), and rabbit anti-α-tubulin (Cell Signaling Technology, Danvers, MA, USA). Secondary antibodies are HRP-conjugated against rat or rabbit (Jackson ImmunoResearch Labs, West Grove, PA, USA). The protein levels were first normalized to α-tubulin, and then normalized to the experimental controls. Densitometry of Western blots was quantified with NIH ImageJ software.

Zhao N, & Zhang J. (2018). Role of alternative splicing of VEGF-A in the development of atherosclerosis. Aging (Albany NY), 10(10), 2695-2708.

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Immunoblot and Confocal Microscopy Analysis

Cited 2 times

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Immunoblot analysis was performed as previously described^{8 (link),35 (link)}, using rabbit anti-TFF3 polyclonal antibody. Mouse anti-β-actin, mouse anti-CDKN1A, mouse anti-BCL2, mouse anti-CDK2, rabbit anti-CDK4, rabbit anti-CCNE1, mouse anti-cSRC, and mouse anti-CCND1 antibody was obtained from Santa Cruz Biotechnology, CA. Rabbit anti-p-cSRC antibody were obtained from Cell Signaling, USA. Rabbit anti-pSTAT3(Tyr705) and mouse anti-STAT3 antibodies were obtained from Abcam, Cambridge, MA. Confocal microscopy scanning was performed as previously described^{51 (link)}. Secondary antibody, Alexa Fluor 488 goat anti-rabbit IgG was purchased from Invitrogen, Singapore. Rhodamine-conjugated phalloidin (Sigma, St Louis, MO) was used to visualize f-actin filaments.

Pandey V., Zhang M., You M., Zhang W., Chen R., Zhang W., Ma L., Wu Z.S., Zhu T., Xu X.Q, & Lobie P.E. (2018). Expression of two non-mutated genetic elements is sufficient to stimulate oncogenic transformation of human mammary epithelial cells. Cell Death & Disease, 9(12), 1147.

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Identifying Cdk4-Expressing Cells in TI

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In this experiment, cdk4 immunoreactivity was newly shown in CA1 cells at 5 days after TI. To confirm cell type of these cells, the sections obtained at 5 days post-TI were processed by double immunofluorescence using rabbit anti-cdk4 (diluted 1:50) (Santa Cruz Biotechnology), mouse anti-glial fibrillary acidic protein (GFAP) (diluted 1:100) (Chemicon International, Temecula, CA, USA) for astrocytes or mouse anti-ionized calcium-binding adapter molecule 1 (Iba-1) (diluted 1:110) (Wako, Osaka, Japan) for microglia by using the method described in Section 2.8. After washing them, the sections were incubated in the mixture of FITC-conjugated goat anti-rabbit IgG (diluted 1:200) (Jackson ImmunoResearch, West Grove, PA, USA) and Cy3-conjugated goat anti-mouse IgG (diluted 1:200) (Jackson ImmunoResearch) for 2 h at room temperature.
To identify the cell type of double immunoreactive cells, the stained sections were observed using confocal microscope (LSM510 META NLO) (Carl Zeiss, Germany).

Lee T.K., Kim D.W., Lee J.C., Park C.W., Sim H., Ahn J.H., Park J.H., Shin M.C., Cho J.H., Lee C.H., Won M.H, & Choi S.Y. (2021). Changes in Cyclin D1, cdk4, and Their Associated Molecules in Ischemic Pyramidal Neurons in Gerbil Hippocampus after Transient Ischemia and Neuroprotective Effects of Ischemic Preconditioning by Keeping the Molecules in the Ischemic Neurons. Biology, 10(8), 719.

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Oxidative Stress Cell Signaling Pathway

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DMEM low glucose, fetal bovine serum, TNF-α, cell culture reagents, and CM-H₂DCFDA were purchased from Invitrogen (San Diego, CA). Primary antibodies, including mouse anti-cyclin D1, rabbit anti-CDK4, mouse anti-cyclin E, rabbit anti-CDK2, rabbit anti-p21, mouse anti-p27, rabbit anti-MMP2, and rabbit anti-MMP9, were purchased from Santa Cruz Biotechnology (CA, USA). Goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Enzo (Farmingdale, USA).

Choi E.S., Yoon J.J., Han B.H., Jeong D.H., Kim H.Y., Ahn Y.M., Eun S.Y., Lee Y.J., Kang D.G, & Lee H.S. (2018). Samul-Tang Regulates Cell Cycle and Migration of Vascular Smooth Muscle Cells against TNF-α Stimulation. Evidence-based Complementary and Alternative Medicine : eCAM, 2018, 1024974.

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Comprehensive Antibody Resource for Cell Signaling

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The following antibodies were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA): rabbit anti-p53, rabbit anti-Bcl2, rabbit anti-Cdk4, rabbit anti-Bcl2, and rabbit anti-p15. Goat anti-p21 was purchased from R&D Systems (Minneapolis, MN), goat anti-Oct4a, and rabbit anti-Msi-1 from Abcam (Cambridge, MA); mouse anti-Hes1 from NovusBio (Centennial, Colorado); rabbit anti-p16, rabbit anti-phospho-PTEN, mouse anti-Rb, rabbit anti-phospho-PDK1, rabbit anti-PTEN, rabbit anti-PD-L1 and mouse anti-Cyclin E from Cell Signaling Technology (Massachusetts, USA). Cell Signaling kindly provided rabbit anti-phospho AKT as part of a sample kit. Mouse anti-PI3K was purchased from Millipore (St. Louis, MO), mouse anti-β-actin from Sigma-Aldrich, and mouse anti rabbit IgG-Alexafluor 647 from Invitrogen (Thermo Fisher Scientific, Waltham, MA).

Nahas G.R., Sherman L.S., Sinha G., El Far M.H., Petryna A., Munoz S.M., Silverio K.A., Shaker M., Neopane P., Mariotti V, & Rameshwar P. (2023). Increased expression of musashi 1 on breast cancer cells has implication to understand dormancy and survival in bone marrow. Aging (Albany NY), 15(9), 3230-3248.

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