MA1-39546

antibody from Invitrogen Antibodies

Targeting: CCND1 BCL1, D11S287E, PRAD1, U21B31

Provider product page for MA1-39546

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [310]
Comments [0]
Validations
Immunocytochemistry [2]
Immunohistochemistry [1]
Other assay [24]

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Product number: MA1-39546 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cyclin D1 Recombinant Rabbit Monoclonal Antibody (SP4)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: This antibody is predicted to react with mouse and rat based on sequence homology. Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is breast carcinoma or mantle cell lymphoma.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Antibody clone number: SP4
Vial size: 1 mL
Concentration: 0.045 mg/mL
Storage: -20°C, Avoid Freeze/Thaw Cycles

CCND1 protein structure - MA1-39546 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Cyclin D1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin D1 Rabbit Monoclonal Antibody (Product # MA1-39546) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Cyclin D1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin D1 Rabbit Monoclonal Antibody (Product # MA1-39546) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 3 Conditional Pax6 expression in Tis21-positive APs increases S-phase in the AP and BP progeny. Dorsolateral telencephalon of tamoxifen-treated E14.5 Tis21 -CreER T2 heterozygous mice electroporated at E13.5 with control ( A , C , D , F , G , I , J , L ) or Pax6-expressing ( B , C , E , F , H , I , K , L ) plasmid (see Fig 2B ). ( A - C ) EdU was administered one hour before sacrifice. ( A , B ) EdU (white) and RFP (red) fluorescence, combined with DAPI staining (blue), on coronal 50-mum vibratome sections. Insets show representative examples of RFP-positive nuclei (outlined by dashed yellow lines) that are either EdU-positive (yellow arrows, see also main panels) or-negative (yellow arrowheads). ( C ) Quantification of EdU- and RFP-positive cells in the cortical wall (total), VZ and SVZ, expressed as percentage of all RFP-positive cells in the cortical wall (200-mum wide area), upon control (Con, white) and Pax6 (black) electroporation. ( D , E ) Proliferating cell nuclear antigen (PCNA) immunofluorescence (white) and RFP fluorescence (red), combined with DAPI staining (blue), on coronal 12-mum cryosections. Insets show representative examples of RFP-positive nuclei (outlined by dashed yellow lines) that show either a punctate immunoreactivity (yellow arrows, see also main panels), indicative of S-phase, or a diffuse immunoreactivity (yellow arrowheads), indicative of G1 or G2. ( F ) Quantification of punctate PCNA-positive nuclei and RFP-positive cells in the cortical w

Supportive validation

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Experimental details: Figure 2 Liver regeneration after partial hepatectomy (PHx) in MICU1 KD and Ctrl liver. ( a ) Representative images of haematoxylin and eosin (H&E)-stained liver tissue and quantification of necrotic area in Ctrl and KD mice before (LL, left lateral lobe) and 30 h after PHx (Phx 30 h; collected from the same mouse for individual comparison). Magnified image displays a typical necrotic area in KD liver 30 h after PHx along with quantification. Scale bars, 200 mum (mean+-s.e.m., n =4 per group). ( b ) Serum levels of alanine aminotransferase (ALT), direct and total bilirubin and triglycerides in Ctrl and KD mice 30 h post PHx, or after sham surgery (mean+-s.e.m., n =3-6 per group). ( c ) BrdU immunostaining and quantification 30 h post PHx in Ctrl and KD mouse liver. Scale bars, 50 mum (mean+-s.e.m., n =4 per group). ( d ) Immunoblotting of Cyclin D1 expression and quantification in Ctrl and KD liver lysates before PHx (LL) and 30 h post Sham operated or PHx (mean+-s.e.m., n =4 per group). Values are mean+-s.e.m., ** P

Supportive validation

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Experimental details: Figure 4 Prevention of mitochondrial Ca 2+ overload rescues liver regeneration in MICU1 KD mice. ( a ) Assessment of Ca 2+ overload-induced mitochondrial PTP opening in Ctrl and KD permeabilized hepatocytes. Extramitochondrial [Ca 2+ ] was recorded during repetitive addition of CaCl 2 boluses (1 muM each) in the absence (Mock) or presence of RuRed (3 muM) or NIM811 (2 muM). Right panel: Ca 2+ load dependence of PTP opening shown as Ca 2+ efflux via PTP (Mock minus NIM811) plotted against the mitochondrial Ca 2+ uptake (RuRed minus Mock), mean traces of triplicates. Representative of n =4 independent experiments. ( b ) Representative images of haematoxylin and eosin (H&E)-stained liver tissue 30 h post PHx showing the absence of necrotic area in both Ctrl and KD liver treated without and with 10 mg kg -1 NIM811 (4 separate mice). Liver necrosis was evident in vehicle (Veh)-treated KD mice. Scale bars, 200 mum. ( c ) BrdU immunostaining and quantification 30 h post PHx in Ctrl and KD mouse liver. Scale bars, 50 mum (mean+-s.e.m., n =5 per group). ( d ) Serum levels of ALT, direct and total bilirubin and triglycerides in Ctrl and KD mice after 30 h of PHx and treated with either vehicle or NIM811. (mean+-s.e.m., n =5 per group). ( e ) Cyclin D1 expression assessed by immunoblotting in Ctrl and KD liver lysates of Ctrl and KD mice treated with NIM811 or vehicle before (LL) and after 30 h PHx (mean+-s.e.m., n =3-4/group). Values are mean+-s.e.m., * P

Supportive validation

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Experimental details: Figure 5 The expression of D-cyclins in mantle cell lymphoma The differential expression of D-cyclin staining in two case examples of mantle cell lymphoma is shown. Case 1 shows intense cyclin D1 and weak cyclin D2 staining. Case 2 shows weak cyclin D1 and strong cyclin D2 staining. Both cases lack staining for cyclin D3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 2 FIGURE Circ-UBAP2 silencing repressed cell proliferation, migration, and invasion, and enhanced apoptosis in vitro. A172 and U251 cells were transfected with si-NC or si-circ-UBAP2. (A) Circ-UBAP2 level was assessed by qRT-PCR. (B) Cell viability was gauged using a CCK-8 assay. (C) Colony formation was evaluated using colony formation assay. (D) Cell apoptosis was detected using flow cytometry. (E, F) Cell migration and invasion were gauged using transwell assay. (G) Cell migration was monitored using wound-healing assay. (H, I) The levels of CyclinD1 and MMP9 by western blot. ** p < 0.01, *** p < 0.001, or **** p < 0.0001

Supportive validation

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Experimental details: 4 FIGURE Circ-UBAP2 modulated glioma cell behaviors via targeting miR-1205 or miR-382. A172 and U251 cells were transfected with si-NC, si-circ-UBAP2, si-circ-UBAP2+anti-miR-NC, si-circ-UBAP2+anti-miR-1205, or si-circ-UBAP2+anti-miR-382. (A, B) miR-1205 and miR-382 expression were assessed using qRT-PCR. (C) Cell viability was gauged using CCK-8 assay. (D) Colony formation was evaluated using colony formation assay. (E) Cell apoptosis was detected using flow cytometry. (F, G) Cell migration and invasion were gauged using transwell assay. (H) Cell migration was monitored using wound-healing assay. (I, J) The levels of CyclinD1 and MMP9 by western blot. * p < 0.05, ** p < 0.01, *** p < 0.001 or **** p < 0.0001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 6 FIGURE miR-1205 overexpression regulated cell behaviors by downregulating GPRC5A. A172 and U251 cells were transfected with miR-NC mimic, miR-1205 mimic, miR-1205 mimic+pcDNA, or miR-1205 mimic+GPRC5A (GPRC5A overexpressing plasmid). (A) GPRC5A protein expression was assessed using western blot. (B) Cell viability was gauged using CCK-8 assay. (C) Colony formation was evaluated using colony formation assay. (D) Cell apoptosis was detected using flow cytometry. (E, F) Cell migration and invasion were gauged using transwell assay. (G) Cell migration was monitored using wound-healing assay. (H, I) The levels of CyclinD1 and MMP9 by western blot. * p < 0.05, *** p < 0.001, or **** p < 0.0001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 7 FIGURE The regulation of miR-382 on cell proliferation, migration, invasion, and apoptosis was mediated by GPRC5A. A172 and U251 cells were transfected with miR-NC mimic, miR-382 mimic, miR-382 mimic+pcDNA, or miR-382 mimic+GPRC5A (GPRC5A overexpressing plasmid). (A) GPRC5A protein expression was assessed using western blot. (B) Cell viability was gauged using CCK-8 assay. (C) Colony formation was evaluated using colony formation assay. (D) Cell apoptosis was detected using flow cytometry. (E, F) Cell migration and invasion were gauged using a transwell assay. (G) Cell migration was monitored using a wound-healing assay. (H, I) The levels of CyclinD1 and MMP9 by western blot. * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 9 FIGURE Circ-UBAP2 silencing repressed tumor growth in vivo. sh-circ-UBAP2-, sh-circ-UBAP2#1-transduced, or sh-NC-infected U251 cells were implanted into the nude mice ( n = 6). (A) Growth curves of the xenograft tissues. (B) Representative pictures and average weight of the tumors tissues. The levels of circ-UBAP2, miR-1205, and miR-382 were measured using qRT-PCR (C), and GPRC5A protein expression (D) and CyclinD1 and MMP9 levels (E) by western blot in the excised tumor tissues. * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Reduced cell proliferation in Nrf1 f/f ;Six3-Cre retina. ( a , b ) BrdU labeling of E13.5 wildtype ( a ) and Nrf1 f/f ;Six3-Cre ( b ) retinal sections. ( c ) The number of BrdU+ cells in wildtype and Nrf1 f/f ;Six3-Cre sections. ( d , e ) E13.5 wildtype and Nrf1 f/f ;Six3-Cre retinal sections labeled with anti-PH3 antibody. ( f ) The number of PH3+ cells in wildtype and Nrf1 f/f ;Six3-Cre retinal sections. ( g , h ) Immunostaining of E13.5 wildtype and Nrf1 f/f ;Six3-Cre retinal sections with anti-Ccnd1 antibody. ( i ) qRT-PCR analysis of Ccnd1 and Gli1 in E13.5 WT and Nrf1 f/f ;Six3-Cre retinas. Scale 50 mum. WT: wildtype

Supportive validation

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Experimental details: Figure 4. CDCA4 affected cell cycle and AKT/mTOR signaling pathway. (a-b) GSEA was utilized to identify the relative signaling pathway associated with high CDCA4 expression. (c-f) The effects of CDCA4-knockdown (c-d) and -overexpression (e-f) on the AKT/mTOR relative markers and Cyclin D1 protein were evaluated using western blot analysis. (d) Quantification of c. (f) Quantification of e. ** p < 0.01 vs . si-NC or vector group.

Supportive validation

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Experimental details: Figure 2 Representation of the final score estimation in the immunoexpression profiling: ( A ) Results of the immunoexpression profiling classified in absent or low staining and moderate or strong staining categories for p16, p21, Bcl-2, Raf kinase inhibitor protein (RKIP), and cyclin-dependent kinase D1 (Cyclin D1); and ( B ) Median size of the appendiceal NENs according to the molecular markers' staining score, (bars with s.e.m., * p < 0.05).

Supportive validation

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Experimental details: Figure 5. Effects of knockdown or overexpression of ADAM29 on proliferation- and motion-related proteins. Western blot assay was adopted to determine the effect of ADAM29 knockdown (a) or overexpression (b) on the level of Cyclin D1, PCNA, MMP9 and Snail. ** p < 0.01 vs. si-con group or vector group.

Supportive validation

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Experimental details: Negative correlation between ecto-enzymes and CCND1 expression in CRC. ( A ) IHC staining of human normal colon (n = 40) and colon adenocarcinoma (n = 64) specimens in a tissue microarray for CCND1 and CD163 (a macrophage marker). ( B ) Relative intensities of CCND1 staining in the lamina propria of a normal and cancerous colon. ** P < .01 (unpaired t test). ( C ) Correlation analysis of NT5E and CCND1 expression in the TCGA colorectal adenocarcinoma dataset. *** P < .001 (Student t test).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Cyclin D1/CDK4 expression is correlated to the presence of KRAS mutation in lung cancer tumours. A. pRb, CDK4 and cyclin D1 immunoperoxidase nuclear staining in lung adenocarcinoma (ADC, upper panels) and in squamous cell carcinoma (SCC, lower panels). CyclinD1 (panel 1, score of 70%; panel 2, score of 40%), CDK4 (panel 3, score of 50%; panel 4, score of 50%), and pRb (panel 5, score of 10%; panel 6, score of 20%) expression. Bars, 100 um (ADC) and 50 um (SCC). B. Distribution of cyclin D1, CDK4, and pRb staining (percentage of positive cells, Y-axis) according to the histological subtype of tumours. Statistical analysis was carried out using Mann-Whitney's U -test. C. Cyclin D1 expression according to KRAS mutation status of tumours. Statistical analysis was performed using Fisher's exact test. Missing data were excluded from the analysis.