Anti-GAPDH抗体- Loading Control
参阅全部 GAPDH 一抗
兔多克隆抗体to GAPDH - Loading Control
Rabbit
From Mar 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. You may also be interested in our alternative recombinant antibody, ab313650.
适用于: IHC-P, WB, ICC/IFmore details
与反应: Mouse, Human
预测可用于: Rat, Chicken, Dog, Saccharomyces cerevisiae, Xenopus laevis, Schizosaccharomyces pombe, African green monkey
Full length native protein (purified) corresponding to Human GAPDH.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 1% BSA, 98.98% PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
浓度
250 µg 浓度为 1 mg/ml
100 µg 浓度为 1 mg/ml
Protein A purified
多克隆
IgG
Abpromise™承诺保证使用ab9485于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| WB | (74) | 1/2500. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa). Some customers have experienced that milk significantly decreases the signal in WB compared to BSA. In-house we use BSA. We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) secondary antibody. Read More |
| ICC/IF | (6) | Use a concentration of 5 µg/ml. We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody. |
Entrez Gene: 374193 Chicken
Entrez Gene: 2597 Human
Entrez Gene: 100042025 Mouse
Entrez Gene: 14433 Mouse
Entrez Gene: 380259 Xenopus laevis
Omim: 138400 Human
SwissProt: P00356 Chicken
SwissProt: P04406 Human
SwissProt: P16858 Mouse
SwissProt: P51469 Xenopus laevis
Unigene: 544577 Human
Unigene: 592355 Human
Unigene: 598320 Human
Unigene: 304088 Mouse
Unigene: 309092 Mouse
Unigene: 317779 Mouse
Unigene: 343110 Mouse
Unigene: 392463 Mouse
Unigene: 392480 Mouse
Unigene: 414470 Mouse
Unigene: 458138 Mouse
Unigene: 458416 Mouse
Unigene: 475698 Mouse
Unigene: 129558 Rat
Unigene: 91450 Rat
Unigene: 995 Xenopus laevis
38 kDa BFA-dependent ADP-ribosylation substrate antibody
aging associated gene 9 protein antibody
Aging-associated gene 9 protein antibody
BARS-38 antibody
cb609 antibody
EC 1.2.1.12 antibody
Epididymis secretory sperm binding protein Li 162eP antibody
G3P_HUMAN antibody
G3PD antibody
G3PDH antibody
GAPD antibody
GAPDH antibody
Glyceraldehyde 3 phosphate dehydrogenase antibody
glyceraldehyde 3-PDH antibody
Glyceraldehyde-3-phosphate dehydrogenase antibody
HEL-S-162eP antibody
KNC-NDS6 antibody
MGC102544 antibody
MGC102546 antibody
MGC103190 antibody
MGC103191 antibody
MGC105239 antibody
MGC127711 antibody
MGC88685 antibody
OCAS, p38 component antibody
OCT1 coactivator in S phase, 38-KD component antibody
peptidyl cysteine S nitrosylase GAPDH antibody
Peptidyl-cysteine S-nitrosylase GAPDH antibody
wu:fb33a10 antibody
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)
All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1/2500 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) secondary antibody at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab9485 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) secondary antibody at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)Image from Wu T et al., PLoS One, 14(4), Fig 3.; doi: 10.1371/journal.pone.0216042. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.
HEK293 cells stably transfected with pINDUCER10-shNF90/NF110 (D2) or pINDUCER10-shNF45 (D5) were treated without or with doxycycline for 96 h, then serum starved for 12 h and treated with PMA (20 ng/mL) for 2 h. Protein lysates (20 µg/lane) were separated by SDS-PAGE and transferred to PVDF membranes.
Loading control: Rabbit polyclonal to GAPDH (ab9485) at 1/1000 dilution.
Secondary antibodies (HRP) were used at 1/10,000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody - Loading Control (ab9485)
IHC image of ab9485 staining GAPDH in human pancreas formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9485, 5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)
All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1 µg/ml
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HEK-293 cell lysate
Lane 5 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 37 kDa
Western blot image using 4-20% Optiblot gel with the Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623).
20ug of Lysate per lane and detection using ab9485 diluted to 1ug/ml.
Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: HepG2 cell lysate.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9485)
ab9485 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab9485 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)This image is courtesy of an anonymous Abreview
All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1/1000 dilution
Lane 1 : Mouse hepatocytes - untreated
Lane 2 : Mouse hepatocytes - treated with LPS (100 ng/mL) for 1 hour
Lane 3 : Mouse hepatocytes - treated with LPS (100 ng/mL) for 12 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-rabbit secondary antibody (HRP) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDa
Exposure time: 1 minute
Primary incubation: 16 hours at 4°C
Blocking: 5% milk for 1 hour at room temperature
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)This image is a courtesy of Anonymous Abreview
All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1/2500 dilution
Lane 1 : Lysate prepared from human Huh-7 cells at 2 µg
Lane 2 : Lysate prepared from human Huh-7 cells at 20 µg
Secondary
All lanes : HRP-conjugated sheep polyclonal to rabbit IgG at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 40 kDawhy is the actual band size different from the predicted?
Exposure time: 5 minutes
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9485)
ab9485 staining GAPDH in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab9485 at 5μg/ml and ab195889 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-GAPDH antibody - Loading Control (ab9485)
Anti-GAPDH antibody - Loading Control (ab9485) at 1/1000 dilution + Mouse Embryonic lung whole tissue lysate at 30 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa
Exposure time: 15 seconds
