Sp pod kit

Manufactured by Solarbio

Sourced in China

The SP-POD Kit is a laboratory equipment product designed for sample preparation. It provides a simple and efficient solution for processing various types of samples. The core function of the SP-POD Kit is to facilitate sample handling and preparation tasks within a laboratory setting.

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Lab products found in correlation

Ab166776, by Abcam (2 mentions) Hematoxylin and eosin staining kit, by Beyotime (2 mentions) Ab219800, by Abcam (1 mentions) Abs50012, by Absin (1 mentions)

Spelling variants of this product

SP Kit (10 citations) POD kit (5 citations)

9 protocols using sp pod kit

Histological and Immunohistochemical Analysis of Gastrocnemius Muscle

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For H&E staining, gastrocnemius muscles were fixed in 4% paraformaldehyde overnight and sent to Servicebio Co., Ltd. (Wuhan, China) for slicing up and H&E staining.
For immunohistochemistry, the sections of gastrocnemius muscle tissues were stained by using an SP-POD kit (SP0041, Solarbio, Beijing, China). The primary antibodies were anti-MYH1 (GTX17458; Genetex, Irvine, CA, USA; 1:400) and anti-MYH7 (S58; DHSB, Iowa City, IA, USA; 1:100).

Zhang J., Cai B., Ma M., Kong S., Zhou Z., Zhang X, & Nie Q. (2022). LncRNA SMARCD3-OT1 Promotes Muscle Hypertrophy and Fast-Twitch Fiber Transformation via Enhancing SMARCD3X4 Expression. International Journal of Molecular Sciences, 23(9), 4510.

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Multiplex Immunohistochemistry for Tissue Analysis

Cited 1 time

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Immunohistochemistry (IHC) staining was performed in tissue sections as described previously (22 (link)). Sections were incubated with goat anti-NEK7 (ab166776, Abcam, Cambridge, MA, USA) and anti-GSDMD (ab219800, Abcam) overnight at 4°C and stained with a second antibody by using an SP-POD Kit according to the manufacturer’s instructions (#SP0041, Solarbio, Beijing, China). Multiplexed IHC (mIHC) was performed using a multiple fluorescent immunohistochemical staining kit (abs50012, Absin, Shanghai, China) according to the manufacturer’s instructions (23 (link)). Sections were observed using an optical microscope (BX53, Olympus, Tokyo, Japan).

Yan Z., Da Q., Li Z., Lin Q., Yi J., Su Y., Yu G., Ren Q., Liu X., Lin Z., Qu J., Yin W, & Liu J. (2022). Inhibition of NEK7 Suppressed Hepatocellular Carcinoma Progression by Mediating Cancer Cell Pyroptosis. Frontiers in Oncology, 12, 812655.

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Histochemical Analysis of Myosin Heavy Chains

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H&E staining was performed using the Hematoxylin and Eosin Staining Kit (Beyotime, Shanghai, China) following the manufacturer’s protocol. Immunohistochemistry was carried out using SP-POD Kit (SP0041, Solarbio, China) with primary antibodies included anti-MYH1(F59, 1:100, DHSB) and anti-MYH7 (S58, 1:300, DHSB).

Cai B., Ma M., Zhang J., Kong S., Zhou Z., Li Z., Abdalla B.A., Xu H., Zhang X., Lawal R.A, & Nie Q. (2022). Long noncoding RNA ZFP36L2-AS functions as a metabolic modulator to regulate muscle development. Cell Death & Disease, 13(4), 389.

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Skeletal Muscle Fiber Typing Analysis

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For H&E staining, gastrocnemius muscle tissues were immersed in 4% paraformaldehyde and then embedded in paraffin and cut into 4-μm-thick transverse sections. Subsequently, the sections were stained with H&E.
Myosin-ATPase staining was carried out using an ATPase staining solution kit (G2380, Solarbio, Beijing, China), following the manufacturer’s instructions. Myosin-ATPase staining was performed at pH 10.4. Under these alkaline conditions, MyHC I isoforms were inactivated while MyHC IIb isoforms were still functional, resulting in addition of black dye to MyHC IIb-positive muscle fibers.
An SP-POD kit (SP0041, Solarbio, Beijing, China) was used for immunohistochemistry as recommended by the supplier. The primary antibodies included anti-MYH1 (F59, 1:100, DHSB) and anti-MYH7 (S58, 1:300, DHSB) and were used for labeling the signals.

Cai B., Li Z., Ma M., Zhang J., Kong S., Abdalla B.A., Xu H., Jebessa E., Zhang X., Lawal R.A, & Nie Q. (2020). Long noncoding RNA SMUL suppresses SMURF2 production-mediated muscle atrophy via nonsense-mediated mRNA decay. Molecular Therapy. Nucleic Acids, 23, 512-526.

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Muscle Fiber Protein Expression Analysis

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Immunofluorescence was performed using anti-MyHC (B103; DHSB, USA; 2.5 mg/mL), and images were captured using a fluorescence microscope (DMi8; Leica, Germany). The area of cells labeled with anti-MyHC was measured and calculated as previously described [20 (link)].
Immunohistochemistry was carried out using an SP-POD Kit (SP0041; Solarbio, China) with primary antibodies including anti-MYH1A (F59, DSHB, 1:100) and anti-MYH7B (S58, DSHB, 1:300). The number of myofibers labeled with anti-MYH1A or anti-MYH7B was calculated.
Hematoxylin and eosin (H&E) staining was performed using muscle tissues embedded in paraffin and cut into 4-mm-thick transverse sections. Subsequently, the sections were stained with H&E.

Ma M., Cai B., Zhou Z., Kong S., Zhang J., Xu H., Zhang X, & Nie Q. (2023). LncRNA-TBP mediates TATA-binding protein recruitment to regulate myogenesis and induce slow-twitch myofibers. Cell Communication and Signaling : CCS, 21, 7.

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Neurofilament Immunostaining in Optic Nerve

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Cryosections of the optic nerve were performed using the same methods as for immunofluorescence. Then fixed sections were treated with 0.1% Triton X-100 and 0.6% hydrogen peroxide. Immunohistochemical staining was performed according to the standard procedure of the SP-POD Kit (Solarbio Life Science, Beijing, China; SP0021) instructions. The primary antibody used in this work is anti–neurofilament heavy peptide (NF-H) (Abcam, ab40796). Three sections of each optic nerve were randomly selected for immunostaining. The image of the proximal segment of the optic nerve at the edge of the injured site was captured by an Olympus optical microscope. The NF-H⁺ area and the tissue area were measured by Fiji software at a fixed threshold. The results of the three sections were averaged to obtain the NF-H⁺ area and the tissue area for each optic nerve. The percentage of NF-H⁺ area was determined by the following formula: (the NF–H⁺ area)/(the tissue area) × 100.

Zhou J., Lin S., Hu Q., Li X., Chen X., Luo L., Ye S., Liu W, & Ye J. (2023). Microglial CD11b Knockout Contributes to Axonal Debris Clearance and Axonal Degradation Attenuation via IGF-1 After Acute Optic Nerve Injury. Investigative Ophthalmology & Visual Science, 64(5), 7.

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Immunohistochemical Analysis of NEK7 in Pancreatic Cancer

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Tissues were sliced to sections of 4 μm. Endogenous peroxidase activity was blocked with methanol containing 0.3% hydrogen peroxidase. Antigen retrieval was performed by boiling in a microwave oven (citrate buffer, pH 6.0), as described before (29 (link)). Sections were incubated with antibody targeting NEK7, goat anti-NEK7(ab166776, Abcam) overnight at 4°C and stained with second antibody by using an SP-POD Kit according to the manufacturer’s instructions (#SP0041, Solarbio, China). Since there were no notable differences in NEK7 staining intensity, we evaluated the ratio of NEK7-positive pancreatic cancer cells. We counted the number of NEK7-positive cells among pancreatic cancer cells in at least 5 fields per section at 200× magnification. Samples were divided into NEK7-positive and NEK7-negative groups; NEK7-positivity was determined when the percent of NEK7-positive pancreatic cancer cells was greater than 5%. We investigated the correlation with NEK7-positivity with survival time, disease-free survival and clinicopathologic factors.

Yan Z., Qu J., Li Z., Yi J., Su Y., Lin Q., Yu G., Lin Z., Yin W., Lu F, & Liu J. (2021). NEK7 Promotes Pancreatic Cancer Progression And Its Expression Is Correlated With Poor Prognosis. Frontiers in Oncology, 11, 705797.

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Muscle Fiber Analysis via Immunofluorescence and IHC

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Immunofluorescence was performed using anti-MyHC (B103; DHSB, USA; 2.5 μg/mL), and images were captured using a fluorescence microscope (DMi8; Leica, Germany). The area of cells labeled with anti-MyHC was measured and calculated as previously described.²⁹
Immunohistochemistry was carried out using an SP-POD Kit (SP0041; Solarbio, China) with primary antibodies including anti-MYH1(F59, DHSB, 1:100) and anti-MYH7 (S58, DHSB, 1:300).
Hematoxylin and eosin (H&E) staining was performed using muscle tissues embedded in paraffin and cut into 4-μm-thick transverse sections. Subsequently, the sections were stained with H&E.

Cai B., Ma M., Zhang J., Wang Z., Kong S., Zhou Z., Lian L., Zhang J., Li J., Wang Y., Li H., Zhang X, & Nie Q. (2021). LncEDCH1 improves mitochondrial function to reduce muscle atrophy by interacting with SERCA2. Molecular Therapy. Nucleic Acids, 27, 319-334.

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Immunohistochemical Analysis of MYH1A and MYH7B in Gastrocnemius Muscle

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An SP-POD kit (SP0041, Solarbio, Beijing, China) was used for immunohistochemistry as recommended by the supplier. The primary antibodies included anti-MYH1A (GTX17485; GeneTex, Irvine, CA, USA; 1:400) and anti-MYH7B (S58; DHSB, Iowa City, IA, USA; 1:100) and were used for labeling the signals.
For HE staining, gastrocnemius muscle tissues were immersed in 4% paraformaldehyde and were then embedded in paraffin and cut into 4 mm-thick transverse sections. Subsequently, the sections were stained using the Hematoxylin and Eosin Staining Kit (C0105S, Beyotime, Beijing, China) according to the manufacturer’s instructions.

Ma M., Cai B., Kong S., Zhou Z., Zhang J., Zhang X, & Nie Q. (2022). PPARGC1A Is a Moderator of Skeletal Muscle Development Regulated by miR-193b-3p. International Journal of Molecular Sciences, 23(17), 9575.

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